Pegasus Laboratories v. US Compounding

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FILED
US DISli{ICT COURT
EASTERN DISTRICT ARKANSAS
IN THE UNITED STATES DISTRICT COURT FOR THE JAN O
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EASTERN DISTRICT OF ARKANSAS " 2 14
WESTERN DIVISION JAMES W. McCORMACK, CLERK
PEGASUS LABORATORIES, INC., )
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By: =::liQ
DEPCLERK
Plaintiff,
Case No. l.\'.\l\ c,\1 \4 ::1'\Jo\
vs.
US COMPOUNDING, INC.,
JURY TRIAL DEMANDED
Defendant.
., ',-:;a assigned to District Judge
,: 111 ::(! 1\,tagistrate Judge - - -----
COMPLAINT AND DEMAND FOR JURY TRIAL
Plaintiff Pegasus Laboratories, Inc., by its undersigned attorneys, for its Complaint
against Defendant US Compounding, Inc. alleges as follows:
THE PARTIES
1. Plaintiff Pegasus Laboratories, Inc. (hereinafter, "Pegasus" or "Plaintiff') is a
corporation organized and existing under the laws of the State of Missouri, with its principal
office located at 8809 Ely St., Pensacola, Florida 32514. Pegasus is a developer, manufacturer,
and marketer of animal health products under the PRN Pharmacal brand name.
2. Pegasus is informed and believes, and on that basis alleges that Defendant US
Compounding, Inc. (hereinafter, "US Compounding" or "Defendant"), is a corporation organized
and existing under the laws of the State of Arkansas, with its principal office located at 1270
Don's Lane, Conway, Arkansas 72032. US Compounding is a pharmacy that engages in
compounding medications, including those sold in the animal pharmaceutical industry.
JURISDICTION AND VENUE
3. This action arises under the patent laws of the United States and is an action for
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patent infringement under 35 U.S.C. 271, et seq.
4. This Court has subject matter jurisdiction under 28 U.S.C. 1331 and 1338(a),
because this action arises under the Patent Act set forth at Title 35 of the U.S. Code.
5. Venue is proper in this District under 28 U.S.C. 1391(b)-(c) and/or 1400(b)
because a substantial part of the events giving rise to the claims occurred in this District and
because US Compounding is a resident of this District.
6. This Court has personal jurisdiction over US Compounding in this action on the
grounds that US Compounding conducts business in the State of Arkansas and has committed
acts of patent infringement in the State of Arkansas.
FACTUAL BACKGROUND
The patents at issue
7. U.S. Patent Number 5,747,476, U.S. Patent Number 6,255,308, and U.S. Patent
Number 6,448,252 (collectively, the "Patents in Suit") are directed generally to compositions and
methods for treating equines afflicted with equine protozoal myeloencephalitis ("EPM") with
therapeutic compositions comprising pyrimethamine and sulfonamide, including sulfadiazine.
8. U.S. Patent Number 5,747,476 ("the '476 Patent") entitled "TREATMENT OF
EQUINE PROTOZOAL MYELOENCEPHALITIS" was duly, validly, and legally issued by the
United States Patent and Trademark Office on May 5, 1998. A true and correct copy of the '476
Patent is attached hereto as Exhibit A and made a part hereof by reference as though fully set
forth herein.
9. United States Patent Number 6,255,308 ("the '308 Patent") entitled
"TREATMENT OF EQUINE PROTOZOAL MYELOENCEPHALITIS" was duly, validly and
legally issued by the United States Patent and Trademark Office on July 3, 2001. A true and
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correct copy of the '308 Patent is attached hereto as Exhibit B and made a part hereof by
reference as though fully set forth herein.
10. United States Patent Number 6,448,252 ("the '252 Patent") entitled
"TREATMENT OF EQUINE PROTOZOAL MYELOENCEPHALITIS" was duly, validly, and
legally issued by the United States Patent and Trademark Office on September 10, 2002. A true
and correct copy of the '252 Patent is attached hereto as Exhibit C and made a part hereof by
reference as though fully set forth herein.
IDEXX Laboratories, Inc.'s ownership of the Patents in Suit
11. IDEXX Laboratories, Inc. ("IDEXX"), having a principal office at One IDEXX
Drive, Westbrook, Maine 04092, has been the sole assignee of each of the Patents in Suit since at
least September, 2004.
12. IDEXX owns the right, title, and interest to the Patents in Suit.
IDEXX's grant of Exclusive License to Animal Health Pharmaceuticals
13. IDEXX, by an Exclusive License Agreement dated July 9, 2010, granted to
Animal Health Pharmaceuticals, LLC ("AHP") having a principal place of business at 1805 Oak
Ridge Circle, Suite 101, P .0. Box 6292, Fairleigh Station, St. Joseph, Missouri 64506, an
exclusive nationwide license to make, have made, use, sell, offer to sell, lease, import, export,
and develop products covered by the Patents in Suit.
14. IDEXX also granted to AHP per this Exclusive License Agreement the sole right
to sublicense the Patents in Suit to alleged infringers.
15. IDEXX further granted to AHP per this Exclusive License Agreement the right to
bring actions for patent infringement against third parties who infringe one or more of the
Patents in Suit.
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AHP's transfer of rights in the Exclusive License Agreement and ReBalance
Antiprotozoal Oral Suspension Product ("ReBalance Product") to Pegasus
16. AHP, by an Asset Transfer Agreement dated September 30, 2010, and with
IDEXX's consent, validly transferred its right, title, and interest in the Exclusive License
Agreement to Pegasus.
17. Thus, Pegasus has the legal right and authority to bring this action and stop
infringement of the Patents in Suit.
18. AHP, by this Asset Transfer Agreement, further transferred to Pegasus its
marketing rights in the ReBalance Product. The ReBalance Product combines sulfadiazine
and pyrimethamine and is used for treatment of horses with EPM caused by sarcocystic neurona.
The ReBalance Product is the subject of the Food and Drug Administration Center for
Veterinary Medicine ("FDA-CVM") approved New Animal Drug Application 141-240 ("NADA
141-240"), a Freedom of Information Summary of which is attached hereto as Exhibit D and
made a part hereof by reference as though fully set forth herein. Pegasus is the current sponsor
ofNADA 141-240.
US Compounding's infringement of the Patents in Suit
19. US Compounding markets and sells in this District and elsewhere, including via
US Compounding's website (www.uscompounding.com), compounded medication product for
treatment of horses with EPM caused by sarcocystic neurona. This compounded medication
product for treatment of EPM is compounded from bulk active sulfadiazine and pyrimethamine
ingredients. This compounded medication infringes at least one claim of each of the Patents in
Suit.
20. Upon information and belief, US Compounding continues to compound, market,
and sell its infringing product for treatment of EPM in the state of Arkansas and elsewhere.
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COUNT I
(Infringement of the '476 Patent)
21. Pegasus incorporates by reference all previous allegations as if set forth herein.
22. The '476 Patent is presumed to be valid.
23. Pegasus has complied with any applicable marking requirements of 35 U.S.C.
287 with respect to the '476 Patent.
24. US Compounding has infringed and is infringing at least one claim of the '476
Patent in violation of35 U.S.C. 271(a) by compounding, manufacturing, marketing, and selling
the infringing product. US Compounding is further infringing at least one claim of the '476
Patent in violation of35 U.S.C. 271(b) by inducing infringement with its infringing product.
25. US Compounding has had actual notice of the Patents in Suit and of Pegasus'
rights in the '476 Patent since at least June 21, 2013. US Compounding's infringement of the
'476 Patent has been and continues to be willful and deliberate.
26. US Compounding's infringement of the '476 Patent has caused great damage to
Pegasus. The amount of these damages is not yet known, but Pegasus has lost profits and
royalties as a result of the infringement and is entitled to damages adequate to compensate it for
the infringement in an amount that is in no event less than a reasonable royalty under 35 U.S.C.
284.
27. Pegasus may also be entitled to recover prejudgment interest, costs, and up to
treble damages under 35 U.S.C. 284.
28. Further, this is an exceptional case under 35 U.S.C. 285 (e.g., due to the willful
nature of US Compounding's infringement) and Pegasus should be awarded its attorneys' fees.
29. As a result of US Compounding's infringement of the '476 Patent, Pegasus has
suffered and continues to suffer irreparable harm and impairment ofthe value of its patent rights,
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and is suffering the violation of its patent rights, all of which will continue unless US
Compounding is permanently enjoined by this Court from infringing the '476 Patent under 35
U.S.C. 283. Pegasus has no adequate remedy at law.
COUNT II
Patent in violation of35 U.S.C. 271(b) by inducing infringement with its infringing product.
the infringement in an amount that is in no event less than a reasonable royalty under 35 U.S.C.
284.
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nature ofUS Compounding's infringement) and Pegasus should be awarded its attorneys' fees.
suffered and continues to suffer irreparable harm and impairment of the value of its patent rights,
COUNT III
Patent in violation of35 U.S.C. 27l(b) by inducing infringement with its infringing product.
the infringement in an amount that is in no event less than a reasonable royalty under 35 U.S. C.
284.
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nature ofUS Compounding's infringement) and Pegasus should be awarded its attorneys' fees.
suffered and continues to suffer irreparable harm and impairment of the value of its patent rights,
PRAYER FOR RELIEF AS TO ALL CLAIMS
WHEREFORE, PlaintiffPegasus Laboratories, Inc. respectfully prays for the entry of the
following orders and judgments against Defendant US Compounding:
(a) Finding that US Compounding infringes, directly or indirectly, each of the Patents in
Suit;
(b) Preliminary and permanent injunctions against US Compounding and its parents,
subsidiaries, divisions, agents, dealers, officers, employees, successors, and assigns,
and all others acting in concert or participation with US Compounding from:
1. making, using, selling, offering to sell, or importing the inventions
claimed in the Patents in Suit; and
u. inducing infringement of the Patents in Suit;
(c) Finding infringement of the Patents in Suit to be willful;
(d) A warding Pegasus such damages, attorneys' fees, costs, prejudgment interest, and
enhanced damages for patent infringement as may be shown by the evidence;
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(e) Finding this to be an exceptional patent infringement case and awarding Pegasus its
reasonable attorneys' fees under 35 U.S.C. 285; and
(f) Awarding Plaintiff such other and further relief as the Court may deem just and
proper.
DEMAND FOR JURY TRIAL
Pursuant to Fed. R. Civ. P. 38, Pegasus demands a trial by jury on all issues so triable.
Dated: January 7, 2014
21136383vl
LATHROP & GAGE LLP
By:
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Laurel E. Stevenson (AR # 2012094)
LATHROP & GAGE LLP
910 East St. Louis Street, Suite 100
Springfield, Missouri 65806
417.877.5917 (Tel)
417.886.9126 (Fax)
lstevenson@lathropgage.com
R. Cameron Garrison
(Pro Hac Vice Application to be
filed)
LATHROP & GAGE LLP
2345 Grand Boulevard, Suite 2200
Kansas City, Missouri 64108-2618
816.292.2000 (Tel)
816.292.2001 (Fax)
cgarrison@lathropgage.com
Attorneys for Plaintiff Pegasus
Laboratories, Inc.
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EXHIBIT A
.,
United States Patent [l9J
Russell et al.
[54] TREATMENT OF EQUINE PROTOZOAL
MYELOENCEPHALITIS
[75] Inventors: Merl Charm Russell. Des Moines.
Iowa; Clan K. Fenger, Lexington. Ky.
[73] Assignee: Mortar & Pestle Veterinary
Pharmacy, Inc .. Des Moines. Iowa
[21] Appl. No.: 683,507
[22] Filed: Jul. 17, 1996
[51] Int. CL
6
....................... A61K 311505; C07D 239/48
[52] U.S. CI .............. - ............................. 514/275; 5441325
[58] Field or Search ..................................... 544!325. 297;
5141275
[56] Refenmces Cited
4.293,547
4,340,609
4,368,193
4,599,416
4,728,641
4,795,639
4,992,444
5,273,970
5,486,.535
5,506,206
U.S. PATENT DOCUMENfS
1011981 l.ewJS et al ............................. 4241180
7/1982 Cbou ....................................... 4241322
1/1983 Argoudelis et a! ..................... 4241180
711986 Kompis ................................... 5441296
3/1988 Tubaro et al ............................. 514/54
1/1989 Burchall et al ......................... 5141249
2/1991 Steveos et al .......................... 5141275
12/1993 McHaroy ................................ 5141157
1/1996 Marr et al ............................... 5141450
4/1996 Kozarich et al .......................... 514/15
arHER PUBUCATIONS
David S. Undsay et al .. "Evalutation of Anti-coccidial
Drugs' Inhibtion of NeosporQ caninum Development in Cell
Cultures." The Journal of Parasitology. Dec. 1989, pp.
990-992.
David S. Lindsay et al.. "Demonstration of Synergistic
Effects of Sulfonamides and Dihydrofolate Reductase/
lbymidylate Synthase Inhibitors Against Neospora caninum
Tachyzoltes in Cultured Cells, and Characterization ot
Mutants Resistant to Pyrimethamine." AJVR. voL 57, Jan.
1996, pp. 68-72.
J.P. Dubey et ai .. "Anticoccidial Activity of 2-SulfamoyJ..-4.
4-Diaminodiphenylsulfone, Sulfadiazine. Pyrimethamine
and Clindamycin io Cats Infected with Toxop!Dsmu
1be Canadian Veterinary Journal. vol. 18. No. J. Mar. 1977.
pp. 51-57.
Daniel Podzamczer et al.. 'Twice-weekly, Maintenance
lberapy with Sulfadiazioe-Pyrirnethamine to Prevent
Recurrent Toxoplasmic Encephalitis in Patients with AIDS."
Annals of Internal Medicine. vol. 123. No. 3, pp. 175-180.
B.T. Nguyen et al .. "Comparative Effects of Cotrim.oxazole
(Trimethoprimsulpharoethoxazole). Pyrimetharoioe-
..;;ulphadlazine and Spiraroycin During Avirulent Infection
with Toxcp/Qsmu gondii (Beverley Strain) in Mice." Br. J.
Pharmac .. vol. 79. 1983. pp. 923-928.
Carol Harris et al .. "In Vitro Assessment of Antimlcroblal
Agents Against Toxoplasmu gondii." The Journal of Infec-
tious Diseases. vol 157, No. 1. Jan. 198S, pp. 14-22.
Harley G. Sheffield et al . "Effect of Pyrimethamine and
Sulfadiazine on lhe Fine Structure IUid Multiplication of
Toxop!Dsma gondii in Cell Cultures." The JoLU'nal of Para-
sitology. vol. 61. No.4. Aug. 1975, pp. 704-712.
11111111111111111
US005747476A
[llJ Patent Number:
{451 Date of Patent:
5,747,476
MayS, 1998
I.G. Mayhew. "Large Animal Neuroloay: A Handbook for
Veterinary Clinicians." Lee & Febiger. Philadelphia. 1989.
pp. 279-285.
Bruce G. Gellin et al.. ''Coccidian Infections in AIDS:
Toxoplasmosis, Cryptosporidiosis. and Isosporiasis." The
Medical Clinics of North America. vol. 76. No, I, Jan. 1992.
pp. 206-234.
Clara K. Fenger et al., "Identification of Opossums (Didel-
phis virginiaoo} as the Putative Definitive Host of Sarco-
cystis Neurona," Journal of Parasitology. vol. 81. No.6. Dec.
1995. pp. 916-919.
"Treatment of Equine Protozoal Myeloencephalitis" by Bar-
bara Brewer Welsch. MA. DVM. et al. 1M Compendium
North American Edltion.Equine. 1991.
"Equine Protozoan Encephalomyelitis" by Till Beech.
V.M.D. Veterinary Medicine/Small Animul Clinician, Dec.
1974. 1562-1566.
"Comparative elfects of cotrimoxazole (trlmelhoprim-sul-
phamethoxazole), pyrimethamine-sulphadiazlne and spira-
mycin during avirulent infection with Toxop/Qsmu goruJii
(Bverely strain) in mice" by B.T. Nguyen et al .. Br. J,
Pharmac. (1983). 79. 923-928.
3615 Eriodicryon, The Merck IDdex, (1989} p. 530.
7997 Pyrimethamine, The MC!'ck Index. (1989) p. 1270.
8874 SulfadifJ7..ine, lbe Merck Index. (1989) p. 1404.
15-Pharmacodynamics, vol. 69. 1968. p. 4749. Sec.
50900p.
Chemical Abstracts, vol. 85. 1976. pp. 46-47, Sec. 72303d.
i-Pharmacodynamics, Vol. 78. 1973. p. 31. Sec. 52708s.
1-Pharmacodynamics, vol. 77. 1972, p. 14. Sec. 109339h.
l-Pharmacodynamics, vol. 92, 1980, p. 63. Sec. 92:1558lp.
Chemical Abstracts, vol. 87. 1977, p. 60. Sec. 87:95742b.
ChemicQl Abstracts, vol. 101. 1984.p. 352. Sec. 101:873391.
ChemicQl AbstrQcts, vol. 99. 1983. p. 20. Sec. 99:133330y.
63-Phannaceutical.J, vol. 96. 1982, p. 373, Sec. 96:40845t.
!-Pharmacology. vol. 123. 1995.p. 37. Sec. 123:102131u.
Chemical Abstracts, vol. 123. 1995, p. 38. Sec. 123:74277a.
Chemical Abstracts. vol. 122, 1995, p. 570. Sec.
122;235089n.
ChemicQ/ Abm'Qcts, vol. 122. 1995, p. 44. Sec, 122:45806w.
1-Pharmacology. vol. 120. 1994, p. 33. Sec. 120:45263a.
!-Pharmacology. vol. 118, 1993. p. 29. Sec. ll8:182836n.
"Pharmacokinetics of intravenously and orally administered
pyrimethamine in horses" by Cyril R. Clarke, BVSc. Ph.D.
et al .. Am. J. Vet. Re.r. vol. 53. No. 12. Dec. 1992.
(List continued on next page.)
Primary Exominer-C. Warren Ivy
Assistant E.wminer-Charanjit S. Aulakh
Attorney. Agent, or Firm-Lowe. Price. LeBlanc & Becker
[57] ABSTRACT
The present invention relates to composltlons and methods
for treating equines. such as hocses. afflicted with equine
protozoal myeloencephalitis or EPM. The therapeutic rom-
positions comprise a combination of pyrimethamine and a
sulfonamide. preferably. sulfadiazine, in the absence of
known therapeutic amounts of trimethoprim.
48 Claims, No Drawings
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5,747,476
Page 2
GrHER PUBLICATIONS
''PhiU'lllllcokilletics, penetration into cerebrospinal Ouid. and
hematologic eifects after multiple oral adminlstratioos or
pyrlmethamlue to horses", by Cyril R. Clarke. BVSc. Pb.D.
et al.. Am. J. Vet. Res., vol. 53. No. 12. Dec. 1992.
"In Vitro Effects of Sulfadiazine and Its Metabolites Alone
and in Combination with Pyrimethamine on Toxcplasrno
gondlf' by Esther Schoondermark-Van De Vco et al .. Anti-
microbial Agents and Chemotherapy, Mar. 1995. vol. 39,
No. 3. pp. 763-765.
"New MJcromethod to Study the Btfect of A!ltimicrobial
Agents on Toxcplasma gondti: Comparison of Sulfadoxine
and Sulfadiazine Jndjvldually and in Combination with
Pyrimethamine and Study of Clindamycio, Metronidazole.
and Clyclosporio A" by D.O. Mack et al .. Antimicrobial
Agents and Chematherapy. Jul. 1984. vol. 26. No. 1. pp.
26-30.
EPM Seminar. The liorse, Nov. 1995, pp. 14-23.
"Equine Protozoal Myelitis Workshop: Summary for the
Horseman" Grayson Jockey Club Resecm:h Foundation
EPM Seminar, Mar. 8. 1996.
',
5,747,476
1
TREATMENT OF EQUINE PROTOZOAL
MYELOENCEPHALITIS
1. FIELD OF THE INVENTION
The present invention relates to compositiollil and melh
ods for treating equines, such as horses,llffi.lcted with equine
protozoal myeloencephalitis or EPM. !PM ls a dehilitating
neurologic disease of equines which can aifect the brain. the
brain stem. spinal cord. or any combination of these three
areas of the equine's central ocrvous system. EPM is caused
by infection by the pl'otozoan parasite Sarwcystit nrurcmu
(recently referred to as Sa falcaru/a). Tbete h no
vaccine or approved animal drug product available fcc
elfectively treating this disease in horses.
2. BACKGROUND OF THE INVEN110N
2
positive serum test Cllllnot be used to ma.ke a diagnosis: such
positive serwn test simply indicates exposure to the parashe.
not necessarily presence of the disease. Cerebrospinal fluid
testing is now believed to be the most useful test to assist in
s the diagnosis of this disease in a live horse.
Cu.rrently available treatment of horses with EPM is
expensive and typically requires a duration of at least ninety
(90) day$. In $ClUe cases. lreatrnent lasts indefinitely, This
CWTent treatment involves the adaptation of tablets intended
H'l for hullllln use. Thus. pyrimethamine tablets are adminis-
tered along with tablets containing a triroethoprlm-
sulfonamide combination. Typically. the two types of tablets
are crushed and placed in suspension for oral administration.
These meWcalions should be administered one hour prior to
lS feeding hay and are accompa.n.ied with frequent. periodic.
veterinary. neurologic examinations during the treatment
period.
Although the symptoms and effects of RPM bavc been l>iscontl
11
uation of therapy is usually based on the admin-
recognized since the 1970's. it was not Until 1991 Ulat Ole lstr.alion of medication Ulirty days beyond the plateau of
protozoan parasite that causes EPM was cul!Ured ftom a 'l1l clinical improvement or disappearance of antibody to the
horse and given the name Sarcocystis neurona. The borse is procozoa from the CSF. Suboptimal dosing or intennittent
an aberrant. dead-end host. as infectious forms of the tltetli!'Y has no proven efficacy.
parasite are not passed from horse to horse or from intected . .
horse to a definitive <ll' true intermediate bost, R-ent Adverse effects of therapy may mclude anemia, abortion.
diardtea and l<IW white blood cell counts. Both medications
investigations indicate that the feces of the opossum (the n for treatment of EPM inhibit folic add metabolism. Unlike
definitive host) are the source of the infection for ht)l$es. ht)l$es. however. the protozoan is unable to utilize pre-
EPM occw-s in much of North America. formed fqlic acid. Supplementation with folic acid or folinic
central one county in Penosylvama. acid (40 rug orally. once a day) and/or brewer's yeast may
and entire states of Ohio and Oregon, have revealed dUll help prevent adverse side effects. It is suggested. however,
approxunately fifty percent (50%) of the borse5 in lbe
30
that foUc acid not be administered at the same time as the
surveyed areas have been exposed to the above-noted pro- pyrimethamine because of competitive inhibition and
tozoan parasite. A positive serum test indicates to abs!Xplion.
the pantsite. not oecessarily the pre6ence of 114 active fonn The gametocytocidal and sporontocidal effects of 2 g
of the disease. The incidence of the adlvc di$CISe is much sulfadialline with 50 mg pyrimethamine in a chloroquine-
lower.
35
resistaDt strain of Plasmodium falciparum is disclosed in
In studies that looked at the disbibution of Sctopo$itive Cbemical Abstracts. Volume 69: 50900p (1968). Primaquine
cases was found that c:ll.riwic fa<:tM may diphosphate, pyrimethamine and sulfadiazine were said to
atfect exposure rates; 1.e .. an frceziag days or a snow causal prophylactic activity against rodent malaria.
very not environment was associated Wl.th a decrease In lhe
40
Pkmnodlum berghei yoelil, as disclosed in Chemical
numbers of horses exposed to the parastte. EPM appears to AbSII'iiCts, Volume 77: 109339h (1972). A three component
have a sporadic .disllibution, al.thougb outbreaks have composition of pyrimethamine. sulfadiazine and
occurred on farms w Kentucky. Ohio. Indiana. Michigan and cycloguanil-H(.'lfor treating rodent malaria is disclosed in
Florida. Cbemical Abslracts. Volume 96: 40845t ( 1982). Similarly.
A horse of any age, breed, cc sex may be affected by EPM.
4
s sulfadiazine sodium has been used to enhance the activities
The disease has occwred in a horse of two IDOIIthS of age, of certain antiiofective drugs against infections caused by
as well as one in its thirties. ID fact. any horse demonstrating pyrimethamine-susceptible or pyrimethamine-resistant
neurologic abnormalities should be considered a candidate strains of P. and P. vivax in owl monkeys. See.
for EPM afll.iction. Cbemlcal Abstracts. Volume 92: 1558lp (1980),
Clinical signs of the condition depend on the location of so There are a number of articles describing the treatment of
the organism within the central nervous system. These signs Toxoplasma gondii with pyrimethamine and sulfadiazine.
include weakness. malposition of a limb, muscle atrophy, but at a ratio that uses a very large amount of sulfadiazine
spinal ataxia. or "wobbling" and/or head tilt with asymmetry relative to pyrimethamine. See. e.g .. Chemical Abstracts.
of the face (e.g., eyelid. ear. or lip). A severely EPM-alfected Volume 7&: 527085 (1973) (I mglkg pyrimethamine and
horse may become recumbent and unable to rise. Lameness
55
100 mglkg sulfadiazine for mice); Oiemical Abstracts.
not traceable to orthopedic disease or any combination of the Volume &5: 72303d ( 1976) (2 mg/kg pyrimethamine and
above signs may occur with EPM. Other unusual signs may 100 mglkg sulfadiazine for cats); Olemical Abstracts, Vol-
also occur. ume 99: 133330y ( 1983); Chemical Abstracts. Volume 101:
In most cases, an atfected horse is bright and alert with a 873391'(1984); Oiemlcal Abstracts. Volume 122: 45806w
normal appetite, although it may be dysphagic (i.e., uoable 60 (1995) (1 mglkg pyrimethamine and 50 mglkg sulfadiazine
to eat) and may act as If It Is choked with feed material !'or monkeys): Olemical Abstracts. Volume 122: 253089o
coming from Its nose. Hematological and biochemical blood (1995). Chemical Abstracts, Volume 123: 10213lu (1995)
values are usually in the normal range. describes the daily administration of 25 mg pyrimethamine
Diagnosis ofEPM is based on clinical signs and on testing and a of 2 g. to prevent toxoplasmic
of the horse's cerebrospinal fluid (CSF). Originally. the 6S encephalitis relapse 10 AIDS patients.
diagnosis was based on the presence of antibodies to Sar- The patent literature includes many descriptions of meth-
oocyslis ncW"ona In serum. tlwugh it is now known that a ods for treating proto:wan-mediated diseases. McHardy, in
,
5,747.476
3
U.S. Pat. No. 5.273.970, states that a protozoal disease.
toxoplasmosis. 11111y be conlrolled to a certain extent using
pyrimethamine together with a sulfonamide. 'This patent
asserts that bequiloprim can be used for the treatment and/or
prophylaxis of protozoal infections in ani:mals, including s
hultlllns. Although the baquilopriro can be used the sole
4
hampering their usefulness in other pathological conditions.
like protozoan-mediated diseases. especially EPM. 'The fact
is that there is currently no approved drug or drug combi-
nation for the treatment of EPM.
3. SUMMARY OF TilE INVENTION
active ingredient. it can be co-administered with a sulfona- Quite surprisingly. it has now been discovered that an
mide. A long list of sulfonamides is provided. most eifec!ive. convenient method of treating EPM is realized by
pre I era bl Y s u If a di aJ.i n e u If a me tho;u zo le the administration to an equine suspected of being attlicted
sulfadimethoxine. sulCadmune. sulfamoxole, or sulfadirni- IO with EPM of therapeutic amounts of pyrimethamine and a
dine. sulfonamide, preferably sulfadiazine. The relative weight
In U.S. Pat. No. 4.599.416. granted to Kompis. a process ratio of pyrimethamine to the sulfonamide may range from
for the preparation of aqueous compositions of sulfonamides about 1:10 to about 1:30. preferably. about 1:15 to about
and sulfonamide potentiators for the treatment of bacterial 1:25. and most preferably about 1:20 in the case of a
infections in humans and ani.ma.ls is described. A long list of 1.:5 composition comprising pyrimetham.IDe and sulfadiazine.
potential sulfonamides is provided. The ''J>otentiators" are It should be pointed out that the compositions of the
described as denoting compounds that increase the antibac- present invention do not contain signilicant amounts of
terlal activity of sulfonamides more than additlvely. An trimethoprim. certainly less than about two-thirds of the
equally long "laundry" list of such polentiators is provided. weight amouat of sulfadiazine present. Preferably. the !hera-
which includes trimethoprim and pyrimethllllline. Other
20
peutic compositions used for the treatment of F.PM are
patents dealing with protozoan parasites Include: U.S. Pat. substantially free of trlmethoprim. most preferably having
No. 4.293,547. granted to Lewis et al. for the treaunent of no trimethoprim at all. S.inilllll'ly. the methods of the present
malaria; U.S. Pat. No. 4.340.609. granted to Chou (various Invention do not rely on the of slgnlllcant amounts
protozoal infestations); U.S. Pat. No. 4.368.193, granted to of trimethoprlm in effecting successful treatment of F.J>M.
Argoudells et al. (malaria); U.S. Pat. No. 4.728.641. granted
15
LJslng substantially the pyrimethamine and a sulfonamide as
to Tubaro et al. (protozoal infections generally); U.S. Pat. the principal active ingredients against the pathologic agent,
No. 4,992.444. granted to Stevens et al. (trypanosomes and namely, the organism Sarcocystis neurona in EPM. Hence.
malaria); and U.S. Pat. No. 5.486.535. granted to Marr et al. the methods of the present invention do not Include the
(To.wpJaslnQ gOIIdii). co-administration of kllown therapeutlc amounts of trlme-
Beech, in Veterinary Medlclne/SmaU Animal Clinic/Qn,
30
thoprim.
pp. 1562-1566 (December 1974) described a condition in In a preferred embodiment of the invention, the atmcted
eight (8) horses with signs of neurological disorder. On the equine. e.g .. a horse. is given a daily dose of pyrimethamine.
assumption that toxoplasmosis was involved. the author which is equivalent to about 0.8-1.2 mg per kg of equine.
suggested that pyrimethamine and sulfadiazine, used sue-
35
most preferably about 1.0 mg per kg. The subject is also
cessfuJly agllinst toxoplaslllll in man. might be useful in given. conCUITently for the greatest convenience, an amount
horses. per day of a sulfonamide. which is equivalent to about 15-30
Welsch, B. 8. recommended the use ol pytimethamine mg per kg of equine, most preferably about 20 rug per kg.
(0.5 mglkg). combilled with a 20 rug/kg two-part mbttlae of Once daily administration of the active illgredients, say
sulfadia.Une (16.7 mgfkg) and lrimethoprim (33 mglkg). to
40
evc::ry morning on an empty stomach. for at least about 3
treat horses suffering from EPM. See. Welsch. B. B . in The months, preferably about 9(}...180 days, is sufficient to treat
Compendium North American EdiTion. Equine. Morris. D. the infection. In some however, the treatment regimen
D. (Ed.) ( 1991) pp. 1599-1602. can last indefinitely. sometimes for the remaining life of the
Two articles by CliU'k et 111.. which appeared in American horse. For ease of administration, the therapeutic composi-
Joumot of Veterinary Research, Volume 53. Number 12,
45
tion may be given orally (that is, by mouth).
pages 2292-2295 and 2296--2299 (December 1992). discuss It should be apparent that an object of the present inven-
the phamlacokinetics of intravenously and orally adminis- lion is the treatmeot of equine protozoal myeloencephalitis
tcred pyrimethamine in horses. The first article. at page or EPM by providing a vetc:riniii)' composition comprising
2292. states that clinical reports indicate the possible value pyrimethamine and a sulfonamide, provided that the com-
of treatment of horses with protozoal encephalomyelitis with
30
position does not also include significant amounts of trime-
pyrlmc:thamine ill combipation with trimethoprim and sui- thopriiiL By "not also include signilicant amounts of trlme-
fomunides. On the other hand. the second article. at page thoprim" means that any trimethoprlm present in the
2299. concludes that the oral administration of 1 mg veterinary composition should not reach any known thera-
pyrlmethamine/kg once a day for 10 days apparently does peutic levels of trimcthoprirn. certainly not reaching an
not present a serious toxicological problem to
5
j amount by weight, which is equivalent to about two-thirds
Hence, despite a great deal ol past and on-going effort, the weight of the active sulfonamide, preferably less than
there remains an unfulfilled need for a treatment for EPM- about one-half and more preferably less than about one-third
aftlicted equines. particuliU'ly horses. which is not only of the sulfonamide. Most preferably. the veterinary compo-
dfective but is also convenient to administa- to maximize sltion of the present Invention (or the Instant method of
compliance apd reduce the emergence of resistant strains. In 60 treatment of EPM) is substantially free of trirnethoprim.
particular, prior compositions for the treatment of EPM Convenient dosage formulations of the present invention
involve three- com pone n 1 mixtures, i 11 c I u di 11 g are also contemplated. including solid and liquid fortns, and
pyrimethamine, sulfadiazine and trimetboprim. Moreover, unit dosage forms comprising about 0.3-0.7 gm
where prior compositions contained pyrimethamine and pyrimethamine and about 6-14 gm sulfonamide, preferably
sulfadlal.ine as the active ingredients, such compositions 6S about 0.5 gm pyrimethamine and about 10 gm sulfonamide.
used very small ratios of pyrimethamine to sulfadiazine These and other objects of the invention will become
limiting their effectiveness to treating malaria. only and apparent to those of ordinary skill in the art. especially after
' ,
.
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5.747,476
5 6
consideration of the following detailed description of the
prefared embodiments.
sal azosulfadimidi oe. sulfa chi oropy ri dazine.
sulfadimethoxine. su!fadoxlne. sulfalene. sulfamerazine.
sulfameter. sulfamethazine. sulfamethomidine.
4, DliTAlLED DESCRIPI10N OF THE sulfamethoxypyridazioe. sulfaperine. sulfaphenazole. sul.
PREFERRED EMBODIMENTS fapyrazine and sulfisomidioe.
The instant invention Involves. in a preferred Still oth<:r su!fonamides known to be useful in vet<:ri.oary
embodiment. the adrninlstration of an oral suspension, con- applications can be adventitiously used, Including
talning pyrimethamine and a sulfonamide. such as phthalylsulfacetamide. phthalylsulfathiazole.
sulfadiazine. designed to overcome the shortcomings of succinylsulfathiazole. sulfabenzamide, sulfacetamide.
cUirently available treatments ofEPM and to provide a more 10 sulfaethidole. sulfaguanidine. su lfamethizole.
effective drug combination for horses and other equines sulfamethoxypyridazlne. sulfanilamide, sulfanilami-
infected with an organism of the genus Sarcocystis. As domethanesulfonjc acid triethanolamine salt, sulfan!tran.
previously mentioned, pyrimethamine may be given in a sulfapyridine. sulfathiazole and sulfisoxazole. The above-
pref<:rred dose of about 1 mglkg equine with a sulfonamide mentioned sulfonamides may be used in place of the pre-
in a dose of about 15 to 30 mg/kg equine. preferably 20 15 ferred sulfadiazine or in addition thereto.
rnglkg. The present compositions may be administered by routes
It has been found that a 30 mL of an oral suspension well known to those skilled in the veterinary and formula-
( such as that described. below) dally on an empty stomach tloa sciences. Theref01:e, although the pyrimethamine and
will provide adequate dosing for the treatment of EPM that sulfadiazine. for c;xample. are conveniently administcted
neither pyrimethamioe nor sulfadiazine can Ileal alone.
20
orally. depending OD the circumstance&. the pl\arnlllc(:utk:a.l
Since RPM is a protozoal infection of the central nervous composition may be administered parentally. topicaliy,
system. the appropriate drug combination must penetrate to intramucosally (e.g .. iotravaginally). or by other routes
the CNS and treat the protozoal infection. known to those skilled i.o this art.
In general. the composition to be administered may com- Compositions suitable for oral admioistrat!on.ln addition
prise about 10-20 gm pyrimethamine and about 150-600
to suspensions. include tablets. capsules. gels, pastes,

gm sulfonamide. preferably 200-400 gm sulfonamide. p<:r boluses. or preparations ia the form of powders. granules. or
liter of composition. The liquid or solid composition may be peHets. Preferred orally administaed compositions include
prepared in unit dosage form depending upon the minimum suspensions and tablets.
size of the equine. Such unit dosage forms comprise a
30
Alternatively. the composition may be administered
relative weight ratio of pyrimethamine to sulfonamide in a parenterally by sub-cutaneous. intramuscular.
range of about 1:15 to about 1:30, pref<:rably about 1:20. intraperitoneal, or intravenous injection. or by .Implantation.
'1)1pically, the unit dosage fonns contain about 0.3-0.7 gmof The composition can be given as an intra.mammary injection
pyrimetham.lne and about 6-14 gm of sulfonamide. most whereby a suspension or solution is introduced into the
0.5 gm pyrimethamine and about 10 gm sulfona-
35
udder.
Jmde. Pharmaceutically acceptable carriers present in the com-
The present invention has been found to successfully positions of the present invention are materials recom-
inhlbit the growth of the organism Sarrocystis neurooo in mended for the putpose of administering the medicament
equines, such mules. ponies aod horses. It has been observed These may be Uquid. sulid. or gaseous materials, which are
that the prefared sulfonamide, su!f11diazine. has resulted In
40
otbetwise lnc::rt or tnedlca11y acceptable and are compatible
better than about a 70% rate of efficacy. In fact. the elfec- wilb the active logtedlenl$. 'The same applies for any added
tlveness of the present compositions aod methods appears to excipieots.
be at least about 80% or 90% of the cases, and probably even For oral administration, fine powd<:rs or granules will
higher. contain diluting agents. for example, calcium carbonate.
In lloe with the foregoin". it is within the contemplation 4S calcium phosphate. mineral carriers, etc .. disbursing aodlor
of the present invention to employ coiDpQsitioos utili.zlng surface active agents. for example, polysorbates, and mAY be
one or more sultonamides udlor pyrimidine derivative in presented In a drench, lD water or in a syrup. in a bolus.
treating ElPM. Examples of other suitable pyrimidine deriva- paste, or iD capsules or sachets in the dry state or in a
tives include. but are not limited to. 2.4-diamino-5- non-aqueous suspension, or in a suspension in wat<:r or
benzylpyrimidlnes substituted in the phenyl ring. such as so syrup. Where desirable or necessary. preserving.
2.4,diamino-S-(3.5-dimethoxy-4-methoxyethoxybenzy)- suspending, thickening or emulsifying agents can be
pyrimid!ne(tetroxoprim) and 2.4-diamino-5-(3.5. induded. If Intended for oral use. a bolus will tx: provided
dimethoxy-4-methylthiobenzyl)-pyrimidine(metioprim). with retention means to inhibit regurgitation. For example.
Still oth<:r useful pyrimidlocs may be 2.4-dilunino-5-( 4- it may be weighed with a heavy density materials such as
bromo-3.5-dimethoxybenzyl)-pyrim.id.i.oe. 2.4-diamino-5- 55 iron or tungsten or the like or may be retained by its shape.
[3.5-diathoxy-4-(pyrrol-1-yl)-benzyl)-pyrimidine. 2.4- for example. by wings which spring after administration.
diamino-5-(3.5-dimethoxy-4-dimelhylarninobenzyl) Boluses may contain disintegrating agems such as maize
pyrimidine. 2,4-diamino-5-(3.4-dimethoxybenzyl)- starch or calcium or sodium methylcelluloses.
pyrimidine (diaveridine) and 2.4-dlamino-5-(2-methyl-4.5- hydroxypropylmethylcellulose, guar based vegetable gums,
dimethoxybenzyl)-pyrimidine. 60 sodium alginates or sodiwn starch glycolates; granulating or
In a similar manner. numerous sulfonamides may be binding agents, such as starch in the form of mucilage. starch
suitably utilized in the present invention, including those derivatives, such as methylcellulose. calcium stearate. talc.
previously disclosed in U.S. Pat. No. 5.273.970. whose gelatin or polyvinylpyrrolidone; and/or lubricating agents.
disclosure Is incorporated in its entirety by reference h<:rein. such as magnesium stearate or stearic acid.
In particular. those sulfona.mides having a dinib'ogen aro- 65 Other compounds which may be included are for
matic ring are especially useful. such as acetyl example, medically inert ingredients. e.g. solid and liquid
sulfamethoxypyrazioe, N-2-formylsulfisomidine. diluents. such as starch or calcium phosphate for tablets.
. '
'
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5.747.476
7
boluses or capsules; olive oil or ethyl oleate for soft cap-
sules; and water or vegetable oil for suspensions or emul-
sions; lubricating agents such as tak or magnesium stearate:
gelling agents such as colloidal clays; thickening agents
such as gum tragaciUith or sodium alginate; dedusting agents
such as liquid paraffin. fixed oils and surfactaots and other
therapeutically acceptable accessmy ingredients, such as
humectants, preservatives. buffers. and anti-oxidants, which
arc useful as carriers in such formulations. When desired,
other medicaments and/or nutrients. such as vitamins or the 10
like. unless contraindicated. may also be included.
It is also to be understood that while the !Xeferred
formulation is administered once a day, it may be given two
or more times a day. dependi.Dg on the circumstances. It
should be understood that while it is convenient to admin- IS
!seer the pyrimidine and sulfonamide concurrendy. they can
be given separately With equal efficacy. However, the othc:r
optimum conditions. such as the administration of the drug
combination on an empty. preferably at least about one hour
before the horse is fed should be observed. 20
A further appreciation of the invention may be gleaned
from the following examples, These specific
e;r.amples are provided for illustration only and are not to be
regarded as restricting the invention in any way.
5. EXAMPLES
VeterinaJ)' compositions effective for the general treat-
ment of EPM are provided. below, in the form of an oral
suspension. The amounts of each component are based on a
liquid suspension having a total volume of about 1 Uter. As
30
mentioned above. a useful dosage, e.g., for a 1,000 pound
horse infected with Sarcocystis neurrma (as evidenced by
the presence of the protozoan in a sample from the subject's
is aboUt 30 mL of the suspension. once
a day. given on an empty stomach in the morning.
35
I Sulfadia.W. B ... , \JSP
z. swt.dlal.iDo Sodium, USP
3. Pyrimelbam.ille USP
4. Sodium lleD:ro<oto, NP
5, Xmt11m pn. NP
6. Aoplll1liJXIt,Nf
1. Sac<:barill, NP
8. y.n,. 111111& (riodictyon

9. ClltU>OJ llovoriQI, NP
10. Polyo:>rbat.e 80, NP
II. Pwified Wt101', USP
166.67 B
166.67 1
16.67.
2.22 a
1.11.
ll.U &
2.78 a
55.S6 mL
5.56 mL
6.67 wL
q,o. to 1,000 mL
Preferably, the composition does n01 contain substantial
amounts of natural sugars. Most preferably. the composition
45
8
3. Mix together the rnaterials prepared in steps 1 and 2 in
a large container and q.s. to one 1.000 liter with the
remaining water.
4. Beat the suspension on a micronizer for live minutes.
being certain said suspension Is evenly mi;r.ed.
5. the final product and refrigerate when nOI in
use. The suspension should be shaken well prior to use.
As further illustrations of the composition of the
invention. the following descriptions of suitable alternative
formulations are provided
Compooonl
I. Sulfodiazin<o Sodium. USP
2 Pyrimelhamlllt, USP
3. SodiUIIl Bcnt.OIo. NF
4. Xonlhan gum, NF
5. NP
7. C..-.mcl llavorina, NF
8. PolyiiOJboote 80, NP
9, Purified water, USP
)33.34 i
33.34
2.22 8
5,0 8
10.0 8
S.56 mL
6.67 wL
'I"' 1,000 wL
The preceding formulation provides an easy to use paste.
Fonnu!atlon c
I. Sulfamotbo,..az.olo, USP
2 Pyrimollwwno, USP
3. Sodilllll llellZOoto, NF
4 Xantban gum. NF
5. NutTasWcel, NP
7. CanuQOI ftowrina, Nl'
8. Polyoorl>au. 80, NP
9. Pllli1ied water, USP
Compouont
I. SulfodiozW<: B-, USP
USP
3. So<lium Belu;ool<:, NP
4, NF
5 Loctoao. NP
333.:!4 B
33.:!4 8
2.22 i
l.ll i
10.0 8
5.!56 Ull.
6.67 mL
q.a. tu 1,000 wL
Per 1000 8
333.34 i
16.67
2.221
10.0 8
638.11 8
is substantially free of natural sugars as some of the sul- 55 ---------------------
fonamides may be sensitive to the presence of oaturally
occuning sugars. Optionally. foUc acid may also be admin-
istered to the subject, either concurrently or at a separate
time. Typically, the subject may receive about 40 mg of folic
acid per 500- to 1000-pound equine.
A stepwise procedure for the preparation of the oral
suspension is provided below:
1. Weigh out the powders and triturate from the smallest
quantity of powder to the largest quantity of powder, so
that all powders are evenly mixed together.
2. In a separate container mill all the liquids IUid add half
the volume of water needed for one liter.
60
65
Fonn!!lauonll
Component
I. Sulfodr:niDo Booc, USP
2. Pyrimetbamine, USP
3. Sodium Bcnzool<:, NP
4. Nutnswcel, NP
5. Lac!OtC, NP
Per 1000 8
300.06 8
!6.67 11
2.22 J
10.0&
671.39 1
The preceding two formulations provide powders, which
can be conveniently divided into individual packets, eacb
containing 30 g of the inventive composition.
9
flonnulauon I'
Compooon!
I. Sul!'aumolo, liSP
2. Pyrimelhamino, USP
3. Sodium Benzoate, NP
4. Xanlhan gum. NP

7. Carzncl Ooavorina, NP
8. l'olyoorbatc NP
9. Purified wafer, USP
formulation G
Per L1ter
333.34 '
n.34,
2.22'
5.0'
15.0.
10.56 mL
6.67 mL
q ... IO 1,000 mL
5.747.476
10
12. The method of claJm lin which treatment is continued
for at least about three months.
13. The method of claim 1 which f\01her comprises
admi.nistering folic acid.
14. The method of claim 13 In whicn the folic acid is
administered daily at a dosage of about 40 mg per 1000-
pound equlne.
15. The method of claim 1 in which the pyrirnetha.millc
and the sulfadiaz.ine are administered in a liquid fonn.
10 Hi. The method of cWrn 1 In which the pyrirnetha.mille
and the sulfadiazine are administered in the fonn of a solid.
17. A vcccrinary composition in unit dosage form foe tne
treatment of equine protozoal myeloencephalitis (BPM)
comprising pyrimetnamine and a sulfonamide In a relative
IS weight ratio of about 1:10 to about 1:30. respectively.
provided that the composition does not also iuclude known
Per Lter tnerapeutic amounts of trirnetnoprim.
18. 1be composition of claim 17 in which the sulfonamide
1. Slllfodimoll!Uxine. 1.1SP 500.10 a is sulfadiazine.
2. """'-tbaminc, USP 16 67
3
, &;;;;;;, BotwJOto, Nl'
2
:
22
: <!O 19. The composition of claim 18 which in the fonn of
4. X.:11han """' NP s.o a an oral suspension.
s. NP 10.0 3 20. The composition of clllirn Ill which comprises about
1. Caremelllavuriog, NP 5.56 wL 10--20 gm pyrimethamine and about 150-600 gm sulfona-
8 Poly110rbatc NF 6.67 wL mlde per liter of composition.
9. 1'\lri&d water, USI' q.e. ro 1,000 mL .., Th u f !aim "0 hi h ri '--
-------------------- 25 ... 1. e compos1 on o c ... w c comp ses avvvt
10--20 gm pyrimethamine and about 200-400 gm sulfona-
Only the preferred embodiments of tne invention and but mlde per liter of composition.
a few examples of Its versatility are shown and described In n. The composition of claim 19 which does not include
the present disclosure. n Is to be understood tnat the inven- substantial amounts of natural sugars.
lion is capable of use in various oilier combinations and 30 ZJ. The composition of claim 17 in which the
environments and is capable of changes and modifications pyrimethamine and the sulfonamJde are present 1n
8
relative
within tne scope of the inventive concept as expressed weight ratio of about 1:15 to about 1:25. respectively.
herein. Foe example. ttle active Ingredients of the contem- :w. The composition of claim 17 in which the
plated veterinary composition can simply be mixed l.o ao
aqueous medium to provide a mixture that can be adminis- pyrimethamine and the sulfonamide are present In a relative
tered to tne affec.:ted equine, usually by mouth. 35 weight ratio of ab<Jut I :20. respectively.
What is clllirned is: 25. The composition of clllirn 17 In which tne uolt dosage
I. A method of treating a Sarcocystis Infection In an comprises about 0.3-0.7 gm pyrimethamine and about 6-14
equine In need of sucb treatment which comprises admin- gm sulfonamide.
istering to the equine tnerapeutically effective amounts of U. The composition of claim 2! In which tne unit dosage
pyrirnetnamine and a sulfonamide, provided that the metnod 40 comprises about 0.5 gm pyrimethamine and about 10 gm
does not also Include the of tne:rapeutlc amounts of trime- sulfonamide.
thoprim. r!. The method of claim 1 wherein the step of admin.is-
2. The method of claim lin which the pyrimethamine and uring therapeutically effective amounts of pyrimethamine
sulfonamide are administered substantially concurrently. and a sulfonamide to an equine includes administering tne
3. The method of claim 1 which comprises administering 45 pyrimethamine and tne snlfonarnide to an equine having an
the pyrimethamine and the sulfonamide orally. empty slomacb.
4. The metbod of claim 3 which comprises the dally .28. The melhod of claim 1 who-cln the step of admlnls-
admlnlstration of the pyrlmetnamlne and the sulfonamide. tering tnerapeutically effective amounts of pyrimethamine
5. The method of claim 1 In which the Step of admlnis- and a sulfonamide to an equine includes administering the
tering comprises administering the pyrimethamine and tne pyrimethamine and the solfonamide to the equine at least
sulfonamide l.o a relative weight ratio of about 1:10 to about one hour before the equine is fed.
method of claim 1 which is effective in at least 2!1. A method of treating equine protozoal myeloenccpha-
about 70% of the cases. Iitls (EPM) comprising administering to an equine iD need of
7. The method of claim 1 In which ttle sulfonamide Is such treatment therapeutis;a1ly effective amounts of
sulfadiazine. pyrimethamine and a sulfonamide. provided tnat the method
8. The method of claim 1 In which tnc solfonamlde is does not also include tne administration of known therapeu-
selected from the group consisting of sulfachloropyridazioc. tic amounts of trlrnethoprirn and who-ein the causative agent
sulfadimethoxine. sulfamerazine. sulfamethazine. of said EPM is other than Tolloplasma gondli.
sulfamethoxypyridaLine. sulfaphenazoJe, sulfapyrazine and 30. The method of claim 2' in which the pyrimethamine
su!fisomidlne. 60 and sulfonamide are administered substantially concur-
9. The method of claim 1 in which the pyrimetna.mine is rently.
administered In a daily dosage of about 1 mglkg of equine. 31. The metnod of claim 29 which comprises the daily
I 0. The metnod of claim 1 in which tne sulfonamide is adml..olstration of the pyrimetna.mine and the sulfonamide.
administered in a daily dosage of about 15 to about30 mglkg 32. The method of claim 29 in which tne step of admin-
of equlne. 6S isterlng comprises administering the pyrimethamine and the
11. The metnod of claim 10 in which the sulfonamide is sulfonamide In a relative weight ratio of about 1: 10 to about
administered in a daily dosage of about 20 mglkg of equine. 1 :30.
.
..
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5.747,476
11
33. The method of claim 29 in which the sulfonamide is
sulfadiazine.
34. The method of claim 2.9 in which the sulfonamide is
selected from the group consisting of sulfachlorpyridazine.
sulfadimethoxine, sulfameruioe. sulfamethuinc.
suuamethoxypyridazine. sulfapheniiZole, sulfapyrazine and
sulfisomid.ioe.
35. The method of cla.im l9 In which the pyrimethamine
is ad.mi.n.istered In a daily dosage of llbout 1 mgfkg of equine.
36. The method of claim 29 In which the sulfonamide is
administered In a daily dosage of about 15 to a boot 30 mglkg
of equine.
37. The method of claim 2.9 io which the step of admin-
istering comprises administering the pyrimethamine and the
sulfonamide to so equine having an empty stomach.
38. The method of claim 37 io which the step of admin-
to the equine at leMI one hour before the equine
Js fed.
39. A method of treating prolo.t.oal myeloencepha-
litis (EPM) coroprisiDg admiulstering to an equine ln need of
such treRtmcnt therapeutically effective amounts of
pyrimethamine and a sulfonamide. provided that the method
does not also include the administration of known therapeu-
tic amounts of trimethoprlm. and provided that the causative
agent of said EPM is other than a Toxoplasma organfSJil.
40. The method of claim 39 in which the pyrimethamine
and sulfonamide are administered substantially concur-
rently.
12
41. The method of claim 39 which comprises the daily
administration of the pyrimethamine and the sulfonamide.
4l. 'Jbe method of claim 39 in which the step of admin-
isterillg comprises administering the pyrimethamine and the
sulfonamide in a relative weight ratio of about 1: lO to about
1:30.
43. The method of claim 39 in which the sulfonamide Is
sulfadiazine.
1
o 44. The method of claim 39 In which the sulfonamide is
selected from the group consisting of sulfachlorpyrldazine,
sulfadimethoxinc. sulfamerazine. sulfamethazine.
sulfamethoxypyridazine. sulfaphenazole. sulfapyrazine and
sulfisomidine.
1S 45. The method of claim 39 in which the pyrimethamine
is administered in a daily dosage of about l :mg/kg of equine.
administered In a daily dosage of about 15 to about 30 mg!kg
of equine.
20
47. The method of claim 39 in which the step of admin-
sulfonamide to an equine having an empty stomach.
48. The method of claim 47 in which the step of admln-
25 istering comprises administering the: pyrimethamine and the
sulfonamide to the equine at least one hour before the equine
ls fed.
* * * * *
' .
: .
EXHIBITB
.
.
(12) United States Patent
Russell et al.
(54) TREATMENT OF F:QUINE PROTOZOAL
MYELOENCEPHALITIS
(75) Inventors: Mt>ri Chnrm Russell, Des lA
(US); Clnra K. hnger, Lexington, KY
(US)
(73) Blue Ridge Pharmuceutlcals, Inc.,
GrccnsboJO, NC (US)
( ) Notice: Subject to 11ny di-;claimer, the terr:J of this
pat.:nt is .:xtcnd<:d or adjusted under 35
U S.C. 154(b) by 0 dl1ys.
This patent is to a terminal dis-
claimer.
(11) Appl. No : 09/069,956
(22) Filed: Apr. 30, 1998
Related U.S. Application Jlutu
( 63) Cominualion of npplication No. mcd ""Jul. 17.
1{)96, now Pat. Nil. 5,747.476.
(51) Int. Cl.
7
. ........... .. A61K 31/505; t\61K 31!:1<
(52) U.S. Cl .......................... 514/256; 514/275; 514/601
(5!1) Field of Search .................................. 51 '1256, 275,
514/601
(56) References Cited
U.S. PATENT DOCUMENTS
3,!'1b,971 9/1975 Mille1 ...................... .., ....... 260/247.5
4.473,548 9/1984 henkel c1 al. ........................ 424;'88
t/J'l87 B<>eckx c1 al ........ - .......... 514.'24:'
l}i52.570 lt!t'JIN Boetb el al. .................... 514:,>.1)
17/l'l9l Mchdwrn t. d;, .. H.u .............
' lt4,'131! ! tl (J9tJ I indct cl al. . .. ..... ....... ... ; 14.'?4:
'1'11.9;\R Hit<N;> l ind 1cr <'I '" .................... '11.'' L'
'./I Ull.1
),fj\)\)l
l.ind.wr t'l 11' '14'
1
1)
5.273,\>711
.
12't'i'l.l Mdla>dy .............. ..... .....
h/1(/{J .... lukami cl al. ................. 514/24'-'
).'16<1.!\.!7 llil Mchlhurn tl al. ................. S 14:242
5, 747.47o 5/!998 Rus.cil el al ...................... 514/275
5,830,893 ll!l'l98 Russed ........................... 514/242
FOREIGN DOCUMENTS
98/43644 10/1998 (WO),
011IER PUBL!Ci\TIONS
Mayhew, ct a!., "Protozoal The Veterinary Clin-
ics of No1th Anwrlco Practice, vol 2, No. 2, p
4:\9 4S9 (Aug. I
W .. "Mtllti!'uc,\l l\curulogic l),,,,<'c >n .\
llor:<.,", Equine Yl'krinHry Scicncl', vol. K. No 1. p
.m2 104 (Apr .. : 'iKK).
I IIIII
l:S006255308Bl
(10) Patent No.:
(45) Date of Patent:
US 6,255,308 Bl
*Jul. 3, 2001
Kobluk, ct (eds.) The Horse Disenses & Clinical
Management, W.B. Company, p. 459-462 and pp.
729-730 ( 1995).
Schoondcrmark et al., "In vitro effect-. of>ulfndiazine and its
alone and in combination with pyrmctharninc on
'Ibxopla;,md Chemical Abstrncts, Vol 122, p. 570,
122.23';0H9(n) (Mar., !995),
Fenger, Clara K., "Update on the Dtagnosis and Treatment
of Equine Protozoal Myclocnccphalits (EPM)," Proceedings
of the D'" Annual Veterinary Medical Fo:-um, American
College of Veterinnry lntl'rnal Medicine, pp. 597-599
(May, 1995).
Andrews, Frank M. et al., "Differentiating ;\feurologic Dis-
cases in the Horse Using Albumin Quotient and IgG Index
Determination," Proceedings of the 13'" Annual Veterinary
l'tHuru," American College of Veterinary Internal
Medicine, (May,
Podzamoer, el al., "Twice-weekly namtcnancc thcraphy
wllh sulfa dia7.ine -pynmcthaminc to prevent recurrent
IOX(>plas>rllL 111 with AIDS", 1-Phor-
mncology, Vol p 17, 121-102111 (u) (Augtsl, llJ'l'i)
Cl.ra K lungvr, i\ppllcntiun of Small Suhunlt Rihn
smnul RNA Glnc ol' Neurunu
toward l>elinentinn of the l)efinitive Host and Scmlugic
Studies (Ph.D. dis.,crlation, l11c Graduate School, Univer-
sity of Kentucky, Lexington (Mur, 1996)) (believed to be
unpublished)
Moore, Bonnie Rush, "Informal Survey of Equine Protowal
Myeloencephalitis," Stale University, Mar., 1996.
Rcrtonc,.loscph .1, "Update ,m Equme Protozoal Mye\oen-
cephnlitJs," FDA Vet., VoL XI, No. Ill, Mayi.Tune (May/June
1996)
f'enger, et a!., "Ep17.ootJc ot' Equine Protozoal Myeloen-
ccphalits on a F.1rm", .Journul ol'theAmcricnn Vctcrimtry
:\1l'dical Associntinn, Vol 210. No 7, p. (Apr.
I ()')7)
cited hy vxam1ner
Primary F.mminer- Kevin E Wedclmgton
(74) Allonwy, Agerll, or Firm-Howrcy Simon Arnold &
White, LLP
(57) ABSTRACT
The present invention relates to and methods
for treating equiGes, such as horses, w.tb equine
protowul myelocm:c:phnhtJS or EPM. the:apeutic com-
a comhumtioo nf pyrirnethaiilinc and n
'ulfonami<k. prcf<:rnhly. 10 the of
k'lllWIJ thl'f.ip< ul:c ot'
29 ('!aim,, 'I/o Drawing"
'
us 6,255,308 131
1
TnEATMENT OF I:QlJINE I'RO'IOZOAL
MYELOENCEPHALITlS
CROSS ltEFERENC'ED TO RELATED
AP!'UC'A'I10N
This app!i,:ation i;. 11 <.:ontwualio;1 of U.S. a,J
1
>1ica.ion Scr
No. 08/683,507 flied Jul 17, 1996 and will into US
Pat. No. 5,747,476 on May 5, 1998.
f1El.D OF THE INVENTION
The present invention relates to compositions and meth-
for treating equines, such as horses, afllicted with equine
protozoal myeloencephalitis or EPM. EPM is n debilitating
neurologk disease of equines which can affect the bra1n, the
brain stem, spinal cord, or any combinatiou of these three
areas of the equine's centrai nervous system. EPM is caused
by infection by the protozoan parasite Sarcocyslis neurona
(recently referred to as SarcocysrL1 Jillcatula). There is no
vaccine or approved animal drug product available for
effectively treating this disease in horses.
BA< 'KGIWUND OF THE
Alihough the and dTccl" of !:PM have hcen
the I rt WI'.' nut until l <J'l\ that th"
proln/.oan that EPM ctdlurcd !'rum <'
horse and given the name neurona. l'hc horse i"
an aberrant, dead-end host, as forms of the
parasite ar not from horse to horse or from mfccted
horse to a dctimtivc or true mtcrmedtatc host. Recent
investigations indicate that the feces of the opossum (tho
definitive host) arc the of the infcctwn for horses.
EPM occurs in much of North America Serologic surveys
conducted in central Kentucky, one county in Pennsylvania,
and the entire states of Ohio and Oregon, have revealed that
approximately ftfly pcrlent (50%) of the horses in the
surveyed an:as have <.:xposed to the above-noted pro-
l<'t.Oan prtrasitc. A positiw serum test indicates <'Xposurc to
2
cominp, fror.1 its nose. Hematological and biochemicai blood
are u>ually in the normal range.
Diagnosis of EPM is based on clinical signs and on testing
of horse\ fluid (CSF). Originally, the
' t!wbno>t'i was based on the presence of antibodie> to Sar-
neurona ir. '<:rum, though it ts now known that a
test ..:annot be lo make a diagnosts;
scrunr t::st exposure to the paras:tc,
.ml nccessanly ut the llu,d
J:; :cstlllg is Dc>W believed to he the most usefultesl to ns.<,ist m
;he of this disease Ill a hve bor5e.
Currently available treatment of horses with EPM is
expellsive and typrcally requires a duration of at least ninety
(90) In some cases, trc&tmcnt lasts indefinitely. 'Ibis
t; current treatment involves the adaptation of tablets intended
for human use. nms, pyrimethamine tablets arc adminis-
tered along with tahlcts containing a trimethoprim-
sulfonamide combination. 'lypically, the two types of tahlets
nre crushed and placed in suspension for oral administration.
7
'; These medications should he administered one hour prior to
fccd111g hay and are nccompanied with freq\Jent, periodic,
veterinary. neurnlogtc cxarmnations during the
penod
DtM:unltnuatJOB or therapy t;, l"lolly based on I he admin-
- or thirty days lbc plntvau o!
clinu.:.ll or ol antibody to thu
protowa !rom the CSF. Suboptimal dosing or intermittent
th..:rapy has no dlicacy.
.'ll Adverse eft'ccts of therapy may include anemia, abortion,
amllow white blood l.uunlb. Both medications
for treatment of EPM inhibit folic acid metabolism. Unlike
horses, howovt:r, the protowan is unal.Jle lo utilize pre-
formed folic acid. Supplementation with folic acid or folinic

(40 mg orally, once a day) and/or may
help prevent adverhe elieets. It is suggested, however,
!hal folk ac1tl not ht: atlminiMcn:tl at lht: same lime as the
pyrimethamine because ol competitive inbibitwn and
the not pr..:M>n<.-c of an form "J
of the diW.ISC. llrc <>I' the ddivc discr,.;c j,_ ,TIJ!t:h
ab,orptinn
l'!tc gl.mclu<:ytociddl and 'Pdr<HJt<x;idal cfi'c..:ts of 2 !\
wtlh 50 mg, pyrirncthamin..: in a
rc"l'tanl ul' fiill'iparun, is clt!-.l:luscd in
( 'lrcrnicul r\!J,u act,, 'vblumc 69: :\!1900p ( 1%8). Primaquine
t.iipltosphalc, and sulfadial'.inc wen: said to
lower.
In stud>c' that lo<'kcd at di'>trihu11un ul
cds<:s gcograph>cally, it was lound thai dimatk ractmh
exposure rates; i e., .m incrc.ase of freezing days or a
very hot enviwnrneot was associated with a decrease u the
numhe1s of horses exposed to the parasite. EPM to
have a sporadic distribut;on, although outbreaks have
occurred on farm; m Kentucky, Ohio, lndia:ta, Michigan and
Florida.
A horse of any age, breed, or sex may be affected by EPM.
The has occurred in a horo;e of two months of age,
as well as one in 1ts thi,l>e,. lu fact, any horst dcmontraung
neurologic -;hould be c<>n,idcred .t
for UP:VI al11>ction.
( 'iinienJ ol c'llflclil>fln :kpend <HI t:w ol
till Wllilin tl:c c'<.:lltr:;lllc fVOW' Wh .. rn "P"
w..:hkn\.,S<.;, tn!dpt.!...,,li(Ht uf a limb, !lHI\l'h: atrnp'l\',
tJ!axw,or dr;d;nl head !.It w.tl "'YJH!lJciJ\'
ol tht (e g, cye.id, c;r lip). Asvvcrely
horse may l'ecome recumbent and to rise Lamcncs'
not traceable to orthopedic disease :Jr any combination <'f the
above s1gns may oclur with l:PM. Other ur.usual may
also occur.
In most an aJJ:'cctcd horse is bright and alert with a
normal appc!Jtc, al!hough it dysphagic (i.e, un.Jblc
to eat) and may act as if it JS chPkcd w.th fcl'd material
J.) prophyladic activity against rodent malaria,
berghei yoelii, as disclos<::d in Chemical
AtJstracts, Volume 77: 109339b ( 1972). A three component
composition of pyrimethamine, sulfadiazine and
cydoguanii-HCI for treating rodent malaria is disclosed in
sc Chemical Abstracts, Volume 96 40845t (1982). Similarly,
sulfadiazine sodium has been used to enhance the activities
of c.'Crtain antiinfcctive dntgs again->t infections caused by
or
of/' fii/crjJ<ii"U/11 and P. t'il'a.\ in owl monkey<;. See,
Cbemll<d i\b,tract;, Volume 92 lS.'i&lp (1980)
Thelt' liH' :> nunrbcr ol arlldcs dc.,crrbing th<> truatrn<::nt <>l
/ilxnp/a\11'11 I{OIIdu wr>lr pyrrmclhamme md
hut at .1 lh.t1 .1 Vt.:''Y c moun I o!'
rdat.vc ll> pynrnc>h.lninc S<.;c, e.g, ( 'bcrnic,d
": Volume}'<'> 52'10ih(l'l73)(1 mg/kgpyrimethamine and 100
mg/kg for mice); Chemical Abstracts, Volume
85: 72303d (1976)(2 mg/kg pynmethamine and 100 mW]<g
sulf.1ciiazinc for cats); Chemical Abstracts, Volume 99:
133330y (1%3); Cher.ti..:al Volume 101: 873391
,,, ( 1984); Chcmilal Abstracts, Volume 122: 45806w (1995) (1
mg/kg pyrimethamine and 50 mg/kg sulfadiazmc for
rnor:kcy>); Chcnu..:al Abstracts, Volume 122 253089n
. '
!
US 6,255,308 Bl
3
(1995). Chem1caJ Volume 123: 10213Ju (1995)
de..-;crihes tht: daily administration of 25 mg
and .1 total ni' 2 g 'ulfadiaLrnc to prevent loxoplasmk
in ;\JI)S pal :cnls.
4
The patent lJicraturc many tk\Cripli<Hh ol rncth- '
m'' l't>f ucating diwa.,c<,. Mdi<rrdy, in
involve three-component mixtures, wcluding
pyrimethumine, sulfadiazine and Moreover,
where prior compo<;tlion'> contained pynmetbnm1nc
sulfaclia/.tne as the
u'cu very hm;rll or pynrnctharr:inc to
licniting their Ill treating malnia only and
hampcnng lhctr w ...,['ulnchh tn other C(r:tlttJlln.'>,
like proto/.oan-mcdiatcd EPM The !'act
that there ih cuncntly Oll approved ll!Ug 01 drug c:omhl-
nation for the treatment of EPM.
lJ S. Pal. No. 5,27},lJ70, lhdt a pnllut.<<ll
toxopla,r:wsis, rnav be controlkd to a certain extent using
pyrimcthami:Je togc:her with a sulfonamide. Tbis patent
asserts t:Jat baqadopruu can be used for the treatment and/or Hl
prophylaxis uf protozoal infecllons in alllmals, induuing
bum;ws. Although the baquiloprim can be used as the sole
active ingredient, it can be co-admiuistcred Wlln a
rmdc. ;\long list of sulfonamides is pmvidt:<l,
prcfurably sull'adia:duc, ;,ulfamelhoxazolc,
1
5
sulfadirndhoxm<:, sulfamoxole, or sulfadimi-
Jme.
SUMMARY OF THE INVENTION
Quite &urpnsmgly, it has now :Jeen discovered that an
t:[ective, t:onvenienl method of treating El'M is realized by
the administration to an equine suspected of being aftlicted
wilh EPM of therapeutic amounts of pyrimethamme and a
sullonamide, preferably sulfadiazine. The relative weight
ratio OJ' pyri111dhaminc to th; ll1a)' roO!o\e f'rorn
uhoul 1:10 to ohou1 1:30, prcfer,tbly, about 1:1 'i to about
ln U.S. Pat. No. 4,'i99,tl It'!, granted to a proces.-.
fr>r the pre par HI ion of aqueous positions ol'
and 'ul!on,lm;tle polcntr.olor' !'or the tn.:almenl of
infe<.:lll>OS in hum:lnS >'llci nnint,Jl'> j, Jonp, or
:H>Ienliilf -;ul!'onHnrdc' i'> f'tovidcd. lltc "putcnti:rlur'>" ;m:
tk:,.cribcd ,1.., compounds t!ldl irtL'n..::l"\1.! 1hc
:erial activlly A more th<lfl add,tiwly. An
equally long "]au:Jdry" bt of <;uch pc>tentintors is provided,
which includes trimethoprim and pyrimethamine. Other
patents dealing w1th include: U.S. Pat.
No 4,293,547, granted to Lewis et al. for the treatment of
malaria; US. Pat. No. 4,340,609, granted to Chou (various
protozoal infestations); U.S. Pat. No. 4,368,193, granted to
Argoudelis cl al. (malaria); U.S. Pat. No. 4,728,641, granted
to Tuharo ct al. (protozoal intecti:>ns generally); U.S. Pat.
No. 4,992,444, granted to Stevens et al. (trypanosomes and
m:.Jaria), anc l;.S Pat. No. 5,486,535, granted to Mnrr c.:t al.
('J'Jxoplavna gondi1).
lkc<.:h, in Vel!'rlflill'\' MedicineiSnw/lllnimal Clinician.
pp. 1 (])e,crrd'<:r )1)71) dl,,r;llcd a l'Pndition ITl
hor""' wrth oi' ncurclugtc,l di,ordcr On the
''''urnption :h..t WI,, lflV{)Ivcd, the author
that pyrimethamine and sull'adin:r.inc,
ccssfully against t<>xoplasma in man, might be t:.scful iu
horses.
Welsch, B. B. recomrnemled t':ie use of pynmethamiuc
(0.5 mg/kg), combined with a 20 mglkg two-part mixture of
sulfadiazine (16. 7 mg./kg) and trimethoprim (3.3 mg/kg), to
treat horses suf:'enng from EPM. Sec, Welsch, B B., in The
Compendium North American 1-.'dirion, quine, Morris, D.
D. (Ed.) (1991) pp. 1599-1602.
Two articles hy Clark ef al., which appeared i:J t\mencan
Jourrwl v( Rnearcl:, Volume 53, Numb.:r 12,
p.>ge., nd 22%-2:'99 (lknmbcr 1 W2),
!he 11:.., n! mlr;l'_.ennu ... ly orn:ly
lcred p,Y!Iff'elh.rn1ifW in hor'c' '!ill' llr'l n!liclc, al

1h:1: cluw . .:d H.porh the valuL
oltrealmcnt of borscs wi:h Wille
pyrimethamine in C<'mb:nation with lnxcmopric:J nnd wl-
fonac:Jidcs. On 1hc other hand, the second article, at page
2299, that the oral admimstration of 1 rng
pyrimethammc/kg once a day for 10 days apparently docs
not prcscot a seri01:s toxkulogJCal problem to horses.
Hence, despite n great deal o: past and on-going effort,
there remains an udt1lfilled need [or n treatment for EPM-
ctfl.1ictcd equines, particularly horses, whkb is not only
cf'cctivc but is also c;(,nvcmenl lo admimstcr to :naximizc
compliance and rcdu..:c the emergence nt' <;train; Jr.
p;nkular, pr.or for tb<: treatment EPM
_,,
1
I :2.1, .1nd mosl prckrably about 1:20 n the or ,,
pyrirncth;ornine and -.ulLHit.rlinc
It he pointed out thai the <.onpositi(>n' (.J' the
present iuYCIIIIL>fl do IIlli \.0.1'.'111 'lgniJk:ant amour:!,'> of
lTI<lWth(Jprim, certainly lcs.'> tl;au about ol tile
25
weight anrouut present. Prderab:y. the thera-
peutic composit1ons used for the treatment of EPM are
substantially free of trimethoprirn, most preferably having
no trimethoprim at all. Similarly, the methods of the prc!>cnt
invention do not rely on the presen(..-e of significant amounts
30
of trimethoprim in effecting successful treatment of EPM,
using the pyrimethamine and a sulfonamide as
the principal active ingredients against the pathologic agent,
namely, the organism Sarcm.:ystis ;Jeuruua in EPM. Hence,
the methods of the present invention do not include the
_,
5
c.:o-admimstration or known amounts of trime-
thoprirn.
In u preferred embodiment ,,f the mwntion, !hu al'fli:lcd
cyuinc, <..p,., d bor,e, '"given,, daily d<NC c>l' pyrimethamine,
whi<.:h i' '-4llJValcnt IL> aboul IUi--1.1 ;-r1g per l:<JUonc,
40 fi1()St prurcrabJy about J.(l rng pl>r kg. rhc 'UbJecl i' dlso
g1vcn, <.:oncurrcntly for the grtaleM c.;onvenicncc, an amount
per day of a sulfonauude, which is equivalent to about15-30
mg per kg of equine, most preferably about 20 mg per kg.
Once daily administration of the active mgredients, say
4
s every morning on an empty stomac:J, for at least about 3
months, preferably about 90-180 days, is sufficient to treat
the infection. In some <:ases, however, the treatment regimen
can last indefinitely, sometimes for the remarning life of the
horse. For case of admini;tratmn, the tbcrapeuti<: composi-
50 lion may be giwn (that ;)., hy mouth).
It be apparent that an obJeCt of the inver:
ticw b th" lrc:tmcnl o ,Jcotowal
or LPM hy providing c\ ising
pvrirncth<ic!linc c111d .t lfldc, provided tha. the ..:orr'-
.. pns11inn dt.H.: .... nul mc..ludt.: :-.ig.mlk'ctiJI arnuu11'o... Dl.ll inH.:-
thoprtnl. By 'not aho include .iHI\lUf :., 1.' trimc-
tbopnm" that any \JJml'!hoprim in the
veterinary composition should not any known :hera-
peulic levels of tnmethoprim, certainly not reaching a:>
60 amount by weight, which is equivalent to about two-thirds
the weight of the active sulfonamide, preferably less tha;t
about one-half and more preferably less than about one-third
of tue sulfouamide. Most preferably, the veterinary cor:Jpo-
sition of the present :nvcntion (or the instant method of
65 treatment of hPM) is substantially free of trimetbopr:m.
Convcnicut dosage formulati;ms of the inwnlw;t
arc also <:onterr.platcc:, !nc<tdmg ;olid :ood ltq111C an:l
US 6,255,308 Bl
5
unit dosage forms comprising about 0.3-0.7 gm
pyrimethamine and about 6-14 gm l>llifooamide, preferably
aoout 0.5 grn pyrirncthaminc wd <trmut 10 grn sulfonamide.
6
dtsclosed t:J U S. Pat. No 5,273,970, whose
is incorporated in its entirety by reference herein.
In particular, th:>'>C a d!nllrogen MO
malic ri11g ar1. u'dul, 'u<.:h a> and other or the lriVCIItion will
apparcnt to ol ortl,nary skill in the .ut, c;,pcciallv i'icr
con-.Kkration ol' the followmg detailed dcM.:np:iou o! the
prd'crrcd cmbodtm<:nt;,.
Dl!Ti\ll.bD DliSC'RIPI'ION or l'HL:
PREFERRED EMBODIMENTS
' nc, N-2-i'onnvbull:-.ornidinc,
' a l :1 " tl.' u i I " d i m 1 d 1 n .: , ;, u ! I " c iJ I o r p y r 1 d a .d u .; ,
'ullHdimdboxtllC, sull'adoxrnc, sulfamerazine,
'u I :a meter, suI fa me th azine,
;ulfarnethoxypyndazme, sulfapenne, sulfaphenazole, sul-
JG fapyrazinc and su lfisomidme.
Still other known to be useful in veterinary
applications can be adventitiously used, wcludmg
ph th al )'Is u If acetamide, p ht h a ly lsu !fat hi a zo lc,
succinylsulfathiazole, sulfabenzamide, sulfacetamide,
The instant invention involves, in a preferred
embodiment, the administration of an oral suspcns10n, con-
taining pyrimethamine and a sulfonamide, sul'h as
sulfadiazine, designed to overcome the of
currently available treatments of EPM and to provide a more
effective drug combination for horses and other equines
infected with an orgamsm of the A>
previously mentioned, pyrirnethaminc may he givtn in ll
pct'crrcd dn-;c or ahout I mg!kg equine with a M;lfonamtdc
in d (\(>.W oJ' ;tllOIII l.'i to :1() Hlg.tkg. c.;quinc, prcl'cr.thly 20
lllgll\:-,.
15 o;u!fdctlndole, sulfaguanidtne, sulfamethizole,
sui famethoxypyndazinc, suI fan 1lamide, sull'amlami-
a1.ad lrieth.trlllhumue sul[anitran,
>.ull'apyr:dnc, sullathJM('k :md wlfisoxat.ole The ahovt-
rnellltonud sulltlfl<trnitk" n.1y be ""cd tn pla<:e ol prl-
,,, lerrecl or tn illhlllic>n tberdn.
II h:" hccn tbumlth.1t a :IOml o! an ural
(su"b as tllat dc:,crihcd, hdnw) \lll empty 1\>m;Kh
wil: pmvidc .H.lcqu"tc !br the treatment n!' !:PM that .,<
neither pyrimethamine nor sulfadiazine can treat :tlonc.
Since EPM is a infectmn of the central nervous
system, the appropriate drug combination must penetrate to
the CNS and treat the protozoal infection.
In general, the composition to be administered may corn-
30
pribc about 10--20 gm and about 150--600
gm s11lfonamidc, preferably 200--400 gm sulfonamide, per
liter of composition. 'The liquid or solid composition may be
prepared in unit do.,agc form depending upon the minimum
size ol' the equine. Such unit tk>sap,e forms n ,.,
Wd!),ht rati<> nJ' pVrtiTI(.!thnlllinc IO in a
flHtgc of .thOIII ! :15 to :tl>out I .\0. pn.d-.,r<ti>ly -dxntl I ::!0.
Typi<.:l ly, the unit dtNtg\. .tl>out 0.:\ 07 gmoi'
.md about 6-14 grn or sull'on,tlnidl-,
preferably 0.5 gm pyrirncthaminc and about 10 gm sulfon<1-
4
,,
m;d<.:.
'Inc present invention bas been found to successfully
inhibit the growth of the organism Sarcocystis neurona in
equines, such as mules, ponies and horses. It has been
observed that the preferred sulfonamide, sulfadiazine, has
45
resulted in bcl!er than about a 70% rate of efficacy. In fact,
the etl'ectiveness of the present compositions and methods
appears to be at least about 80% or 90% of lbc cases, and
probdbly even higher.
In lDc with the foregoing, rt i;, within the conlcrnpblmn ,.,
ol' ihL inwnti(n to employ C<Hnpthition'> uti:t/.in)!.
or.c or more suJonarnid<.' and/:>r pyrintidmc dcriv<tivc in
Ire Ll'M. ui' t>lh..:r '-Uitahlc pyrimidi11c denva-
it:Liudc, but <.rc nt llrnit<:d tn, ?.,.1-dtannrh>-S-
'Jh-.tttutcd in th<. phenyl ring, 'ULh a"
:::> ,4,dian inu-5-( :1,5 d1 mcthoxy-4-mc thoxyc thoxybe nzy )
pyrim id iue( \e twxo pn m) a ud 2 ,4-di arn i no-5-( 3,5,
d :me; ho x y -4 -met hy ltbioben zy I )p yrirnid me( me tiop r int),
Still other u,;cful pyti:mdmes may be 2.4-diamino-5-(4-
bror:I0-3,5-diructhoxybcnzyl)-pyrimidine, 2,4-diammo-5- 6:,
[3,5-Jia thoxy -4-(pyrrol-1-yl)-benzyl}-pyrim idine, 2,4-
dia mino-5 -(3,5 -dt me thoxy-4- dimethyl am 1 nobcn7y 1)-
l'hc J"C<;i.:r:l TidY h<.. udrnHI!sh..:rl.:d hy otlll'.'t
we!! ktl11Wn :t tl:o,c '>kil!t..d in the vei\OfLJHlC) ;md forrnuln-
llon 't'tcnce" Th"n.:i'm<., althOII['.h the pyrirncth<trnwc Hncl
i'or example, ar" conveniently administered
orally, clcpcndwg on the circumstances, the pbarmaceuticnl
composition may be administered parentally, topically,
intramucosally (c g., iotravaginally), or by other routes
known to those skilled in this art.
Compositions suiwble for oral administration, in addition
to suspensions, include tablets, capsules, gels, pastes,
boluses, or preparation> in the form of powder<;, gmnules, or
pellets. Prcl'cned orally adm1mstcr"d compositions- ioch1de
'1tspcn,ions and tablet<;
the rnay bt
parcn!crall) by 'lth-lutallcou,,
inll;!pt:fiHHJC;il, or itll!i\Vl:rtOW tiJC!.'IIlHl, ur by lfllpian:uth)IL
jl,l: Clii'Tlj1U...,li1011Ull1 ht: ).!,!VI..?II ;l.<..,l\1' iojcrlion
wht:rl'i>y 11 '-U.,pcn;,.on 01 ,o:ttton '' intmclu""d inln the
uclclcr
l'harmae<:Utlt:dlly awcptablc \.'ilflli:TS in tbc corn-
positions of the prescn: invention are matenals recom-
mended for the purpose of administermg the medicament.
These may be liquid, solid, or gaseous materials, which are
otherwise inert or medically act.'eplable and arc compatible
with the .tcl!Ve ingredients. The applies for any added
excipients.
For oral adrnintstrahon. fine powders or gram1les will
contain (,l;lulmg agents, for example, calciu:n carbonate,
caki111:l p!JC,ph.1tc, carrie;, etc., ci!sbursing amlior
ucltve kr tX<m;1lc, polyorb.tte,, aod m.ty be
prc,.;ntctl 1n ,< drench. lfl w.il(.; Pr m " iu
,., 111 '-"l''>l.lc, tr 1'1 It" dry '\Hie w 10 a
... tH !!I 1 HI w.1tcr
')'fUj1 or JHCS<.rVtllg,
or CJtl be
im:lucled. If ntenckd for oral L!-<C, a hol% will he provided
witt- retentinn means to inhtbit For ex:unple,
it may be weighed with 3 heavy density ma:enals such
iro:1 or 1\Jngstcn ::1r the like or may be retained by its shape,
for cll.arnpk, by wing> which spring after admmi>tratlon.
Boluses may contain disinkgrating agents such as maize
starch or calcium or sot'iurn
hydroxyprnpylrnetbylccllulose. guar based vegetable gums,
PY rimidi ne, 2,4-dia mino-S -(3,4-d imcthoxybe nzyl)-
pytinmliuc and 2,4-dtamiuo-S-(2-mc;byi-4,S-
dlnwtr.oxybcrll,yl)-pynn:idinc.
lu a manner, IJUrncrous he
'ullably 111 the pi..:scnt mvcntion, th"c
,,,, algio<ttcs or sodium g:ycolatcs; gra:JUlatmg or
htr:clng ag.c:1h, .. h .ts n !hL of
'-tllcn ,1.., Th.thyh:Ll!uln..,'-'. l,,:ciun o;!car.:Hc.
us 6,255,308 ill
7
gelatin or polyvmylpyrrolidonc; <tndlt>i
such ao; stl.!aratc or dl'!O
Other l:OrHFl11111<1' wh11.:h !'lay he' 11-cludcll J,>J
example, mcdkally II"''' c '"lid .a1d lllJllld
dilucnh, 'uch a' '''m:h ur uilu.;m pho,ph11h: (;,,
holus..:s t>f L'.ll''ulus, ol1vc tHI or cll:yl nk111e lor '"II c:op-
-;uJcs; dOd Wd\cr or vcgctahk oil ror or emul-
sions; h1brka1ing agtmls "ud1 asIa:..: or magne,ltlm >leilf.llc,
gelling ngmts su"h a> colloidal tbi,kening agents
such as gum tragacanth or wdium algma1e; dedusting agents
10
such as hquid paratlin, fi:...:d oils and surfnctanls and other
therapeutically acceptable accessory ingrcdtLrHs, sm:h as
humcctanl,, buffers, and anti-oxidants, which
arc useful as carriers in such formulations. When des1red,
other medicaments and/or nutrients, as vitamir1<> or tt:c '
5
like, unless contraindicated, may also be included
II is 10 1>.: umkr,tootl that wnik prclc'l cd
lormul,>lion I'> mlnHill'lcrcd ,,,_._a d.>y, il r,w he >v.<>
or more l111es .t day, 011 l'rCulll..,ldiH.:L' ll 'L
he umlcr,lood llio!l v.hlic 11 '' Lonvc"'"'ll lo .>dmin-
''ler the pymniclinc nnd sullonam>cie they <:illl
he givt:u scparalcly with ,qual cllkacy. llLlWcwr, the uth<:r
optim\lm conditions, such n the aclmmio;tn>IIOO oi lilt: drug
combination on :m empty, preferably at least oboul O;JC hom
25
before the horse 11> feel should be observe-d
A further appreciation of the invention may be gleaned
[rom the following specific examples. specific
examples arc provicletl fm illustration only anc! are nol to be
regarded dS re:;trictiug the inveotwn 111 any w y. .<o
EXAMI'LJ:,S
o.:<>mpo;.ilron' <.:ll'ccliv" f1>r tile trc.ll-
ol .nc fli(Vllk<i, b<:low, m tl>c IL>IIll ,,f <Ill LHal ,,
..;u:-.pt:ll"-lon. The .... of C'-JI.!h umpon\.'nt an: ,)fl d
liqu1d ,u,p<:H:-iun ,1 total volume u> ,,i>oul I hlcL
nwntrom;d above, a usvful e.g., lor a l,OOOpuuml
in fcc ted with ucJ:moa (as ev Jclem:cd by the
of thc pwtolo<Hl in d t'mm tbc ;.ubject Ml
fluid) is about 30 mL of the cnce
a day, giwo on an empty stomach ;n the: morning.
.....foJ.nulati>'' A
\)ut,ldl.itllll' u;,..,<.-, l:sr-
Sul ,HJintiiW 1'111 l '\[l
Py1 1111.'1,\lro k . l ....,p
4 'lodilll" h I'. I
.:-. x .. !' )d l guw. 1\.1
J\;,p.q!.'Jllt:, 'll'
;-,.1...lh.>.tu, Nl
Vtrhi1 -.i'lnld ..

9 < 'ltlmd n.t,'Oll e;. r--.1
10 l'o!yso hr1l1. ,-<II, 'i
Jl t'nd . ..:<J U'3F
Pu
'I'
t._.
l.lt:l
Fo\' (/
1:11
'"
jll
'
"
"'" .<:.L
,.,
:'"!: ml
b 07 mL
l,t;OtJ mL
4'
:-n
,,
Prd'tr:tbly, the nut conlaill <>IJ
amr>unh ll! naturul prcf'nahly, the Ct>mposrliuc
l'> t'ree of na11.nd 'o\l!l,:l'S '"' ":llle ul lhc sd-
1\mnmtth.:::..; rnay lH: \CI':-;ii!Vl ttl' o llPlllrnl!)'
Ol.'i.'Umng Option:tlly, an'' may al"' t1e ->d:>Jlll-
:o thL: .;(Jncurn.:nllv 11! ,t ntt..
ti:11C Typiud!v, me \\!Dll.A ..'l mnv rcu:!Vc it\HWI -til tl. J'D\.;.
uc:Jcl per )00- to l!J(l().pou>d
8
A 'lcpw ise pro<.:eJure l'or the prcpdfdlion ,,f the oral
ll'>!nn provided bclnw:
Wc!)!,h nul lire pow(\er' ,1nd lfllllf.tle l'rorn the ,rn,>IIC'ol
qu;, nlity ol' po'Wdcr to [t,e of powder, Ml
that ,dl pt>wdcJ;, evenly m1xcd togctbcr.
.2 lu a ..:unl,uncr nux all thc liquids anti add half
the volume of Wftter needed for one liter
3. Mix together the matenals prepared in steps I and 2 w
a luge container antl 4" to one 1,000 hter with the
remammg water
4. Heat thv ;uspcnsion on a micronizcr for live minutes,
bting certain said suspen!>ion is 1:vcnly mixed
5. Pl!d,age lbt: prodult dOd rdnger<>tt: wh<>n not in
u,e, The su,pensi,>n be shaken well pritll to uSt.
A' l'urthcr of the composition ,,r the
urvcnliun, the J(,l!owing dc,l'riptinuh D! hUilal>le a!lcmahvc
i'<>IITII!idlltllh iliL pn>v:ded.
h ..mnu !ntion B
Compo11ent.
l. Sutfadiarine Sod.Jum, U ..
2. l)yrimdhamne, USP
3. Sodium lletu;oate, NF
4, gum, Nl'
5 N1ttm'Oweet, 1\'1
7, l tlavoling
1
'Nl
3 so. NF
9, Pl,nht..d W;lll:r, USP
('omj)onenl
1 .. Std(amelhoxazole, lJSP
2 Pynmelh.amine, USP
3. Sod tum Bel:l.i..Cate, !\. E
4 Xnnthon gum,
5 NlltJnr,weet, NF
Caromcl f\.wm tng .. '\ll
Nl
), Put1f1ed w.llel, liSP
!\;..,._, l
1
Pvr 1t1h l1l1lll,
{, SO.JlUIP /hrvwttc,
4, t'Wlid')WC:L(, '\J
Lacto\t::, r-.:r
:::hilt.nJv:>.J \{,. l 'il'
- P\ tdl.eLhcnuH l'S!'
.:),'d!': n he '\;["
t'-.'1, :-..1
-., : ,,._1 ... ,. '\ll
Per LLter
333.34 g
3.L\4 g
2.22 g
5.0 g
lO.U g
5.56 mL
b.(i7 mL
ll .._ ltl ntl
Per
313.}4 &
33 31 g
2.'27 g
Ill S
D.Cg
'i.5ri mL
L61 ntl
(j b ttl ,01.;1:; l"'ll
.. q J,
( {> '
J' (..
Jt) ')
ll g
].00.05 g
lt; 67 g
2
j)(' l"-
:)7 !
p co.:ed.ng lwt. jllOV.dt puwdcf'>
\.dr! i)t' lll il\ly d.\ Hied 111:0 ,mlivH\Ud' Ltii.:h
(,nn!d!fltng lll !l (,f u)mpo:-.itiu:-L
US 6,255,308 Bl
J v.;r
2, Py!trelhl!T'JOI.... I ..;p
3 SodilHH Bervol!te, Nr
-1. Xanhan Ni"
5. Nf
7 C:
1
ra-:nel J'\nvonng, NF
80, NF
..) PtH wa:er. USP
9
F01 m1.1latlon G
i. Sulf3d!m<too<im:, 1 'W
' PynmolhnUline. l 'SP
.l, S.1d1um Bcmwnl<. 1>\1'
>1. XnIIMn
i"1!\HHl:r."H.I,
1
t''11M1I!..'ltl,n.l'llt:,N}
?<> Pn:, ... ,,.h,,tt' -.fll "'t
li.lhiKJ 1. l
333 _q
J4 g
2 22 1:'!
S,l) g
15,0 g
10.55 n1L
6,67 mt
q s :o l,COO n:L
'() I 1 g
0.!)/ g
... ..,, '
,!,,!
Only the prdenecl emhodiml'nts of the invention and btl!
a lew of are :,howo and dc:-.cribed 10
the pn:scnt d!,dosure. It is to he that tht: inven-
tion is capable of use in various other combina\lons and
environments and is capable of ch,mges and modifications
within the scope of the inventiw concept as expressed
herein_ For example, the active ingrcdtcnts of the contem-
plated veterinary composition .::an simply be mixed in an
aqueous to l''ovicl<.' a mtxturc that can l:x: adminis-
tered to the alicctcd equim, U<;Ually by mml\h,
What dnim<!d is
l. A method ol treating, "quinc ,,;otoYmli myclocnccph.t-
(lol'M) .;ompri,mg to an equine
10
9, The methut! of claim I which "' Ill at lea't
about 70Cfc ol the ca-;c'
II) The rnl'thod of' daim 1 in whtch the ,uJonarnide ''
o;ul!atllat.lllc
lL The method of daim I in whtcb the '>ulfonamidc is
frcnu the group con,istmg o[
sulfatlimcthox inc, sulfamerazine, sulfametb azine,
sulfamethoxypyridazmc, sulfaphenawle, sdfapyrazmc and
sulfisomidine.
10 12. The rno:thod of claim 1 in which the pyrimethamine is
administered in a daily dosage of about 1 rng/kg of eqmne,
13. The method of claim 1 in winch the sulfonanude is
admmistcred in a daily dosage of about 15 to about 30 mg/kg
of equine.
t> 14. The rm:tbod or <:!aim 13 in whtch the sulfonarnid.; ''
adiHinistered in a daily doagc of about 20 ntgikg uf clJUHtc .
15. The rrH.:Ihod ol d:11rn l <n wh<.:h tre.1tmcnt1' contmuctl
lor at Jc.1't ahoul three rnunlh'
16 Thl' lflctlHd .. r claiJlt I whid: !IJrtkr
.ldtollll"'lcl fnlu.: <11..1d
\7, method ol cla1m 16 m wh1ch the tol:c aqd ,,.
dally 111 a dn,agc of about 40 rng per 1000-
pound cqu1nc,
18 The method of claim 1 in which the pyrimethamine
zs and the sulfadiazine are administered in a liquid form.
19, 'Ibe method of claim 1 in which the pyrimethamine
and the sulfadiazine arc administered in the form of a sohd.
20. A veterinary composition in unit dosage form for the
treatment of equine protozoal myelol.!nccphnhlls (EP:Vt)
JO pyrimethamine and a sulfonamide in a relative
weight rauo ot' ahout 11 0 \o about I 30,
provided that il' the t' "ulladwtmc the
docs not alo;o include nn arnounl or trmtethopf!fn
Whit:h IS 20 percent i>y weight oJ' the HfTlL>llfll or 'ulf,I-
J_, diat.inc prc<;cnt in the ct>rnpo,i:ion
ot' 't lkr<:Ji'l frwn LI'M <IID<>llllh of
pyrimcthamm.: ami a sullomtmidc, provided that il th.:
sulfonamide '>-ulfadtalmc, an amount of trimethoprim
not abo administered, wh1ch nrnount of trimcthoprim is 20
percent by weight or tbc amount of the admin-
4
o
!Me red
21, The c'<Hnpositionol diiim 211 in wlw:h the 'ullonamidc
i., sulfadtazinc_
22, The composition of claim 21 whtch is in the t'Ol m ol
an oral .'>Uspension.
23. The composition of claim 22 which compnses about
l0-20 gm pyrintethaminc and about 150-600 grn sulfona-
mide per liter of composition.
2 The method of claim 1m the pynmcthamine ;tnd
,ulfouamide are adrniuistered "ubstantially coacurrcnt!y.
3, The method of claim l whtch does not include the
.:o-aclm,mstratwn of tnmcthoprim.
4 metbvcl cl,tim l WhiCh COlllpfies aclmini:;lering
pyn and tit..: '"ally
5 l'be mdbm' ut dann 4 wl:ic:h dRil}
u[ th: pyrimdhammc and th..:
6. r!w fi!CtbLlJ lll <'IHif':l l Which atlllllfi,;,\Crtllg
a thcr;,peuiJually ctlo;;dtve <llll<>llll. nl a nmpu"lio.J cunJ-
.md the ,,ullonanmk, JHllVJd<.:d
lli.it i! I!Jc 'u!tOil<lfTll<IC ,, 'ull:\dtaLIII<' lb.; ..:UfiiJ'tl'ltllHJ due'
fl<t .1ho l'tcitJ<!<! an amount ol trirnethoprirn whKh "' 20
percent by wc1ght L>i'thc ot ,LJiadHtzme prt:""nt
'" th
7 The method or daim 6 lin.: cumvositilln compn:-.c" the
the in a relative weight
rn:io of about 1: I 0 to about 1:30
!! !'he of da1rn l which inhibit<; the growth ot an
24. The composition of claim 23 which <.'Ompriscs about
10-20 gm pyrimethamine and about 200--400 gm sulfona-
-15 tmdc liter of composition.
25 The compositllJn of d.uru 22 which elves not in-:lml<.:
'>Ub,lantl<ll uf O<llural !>Ut-;MS,
26 Till.! <.'Lll'1Jlll>lllllll ur claJIJI 211 Ill wb<dl
pyrum:th:un1uc anti the "ulfnllrllll!dc 111 a <cialtv'
''l 1<1tio ol ! IS tu - 25, :''P''''tlv,,y
27. Ttw ol' <.l<lllll 20 Ill wtncb
pynmcthamto<.: .md th<.: sullun.Hllllk <.rc pr<:>.Cfl. 10 a rciHtiV<'
W"1ght r.ltHl ol ,tbtJLI \ ,20,
211. fh oi <:b,-n 20 tn wtm:b the unit
'' comprises about 0 _1 0. 7 gm and :ohout 6-!4
grn
29 The composition of cla!m 211m which the t;nit dosagt
compnses about 0 5 gm pynmethamme <tnd ahout 10 gm
:;ulforwmide,
I '
' '
' .
EXHIBIT C
',
(12) United States Patent
Russell et al.
(54) TREATMENT OF EQUINE PROTOZOAL
:\-1YELOENCEPHALITIS
(75) Inventors: Mer! Chnrm Russell, Des Moines, !A
(US); Clara K. Fenger, Lexington, KY
(US)
(7'>) Assignee: IDEXX Plmnnaceutlcals, Im:.,
NC (U.S)
( ) Notice Subject to disclaimer, the term or this
ptcn! extended or under :IS
US(' 154(h) hy 0 dnys.
Th1s pte nt is subject to a terminal dis-
cla!mcr.
(21) Appl. No.: 09/685,943
Filed. Oct. tO, 2000
Related U.S. Applicutlon llata
( 63) Conhnuation of upplicution No. 09/0&'1,956, !ileu on Ap1.
30, 1998, now Pat. Nc. whtch is a contuJuation
of npplicn!Jon No. ORI6R3.507. filed on Jul. 17, J'J<Jo, now
Pal. No.
t'il) Int. Cl." A61K 31/505; A61K 31i18
(5'2) U.S. Cl ................. 514/256; 514/258, 514/275;
514/601
(58) Field of ................................ 5141256, 275,
514/601, 258
(56) References Cited
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l .. li>ll,1''-' A et al. ......... 424:180
i\ 9il'IH4 I'Jcnkrl C! .tl ............. 424iH8
A 7tt<)Hu ........ , ...... , .. , 544,"19()
1.7Jfi,h<ll A VJ9i!H TulHIIO ct ,,J . ... ... 'il4i'"'
A X:1
1
Jt!H cl al. ... :d4;242
'.79':"1 1\ lii%\1 lllllehall cl a ....... 'i14.?4'J
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'i,27J,'J70 A 12!1\:93 Me Hardy ............. 514/157
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5.464,H37 ,\ 11/1<;95 Mehlhorn et al. .......... '14/242
A 1/199b Mall ct al. ............. 514,l.''
5,5tJti . .il!o 1\ cl l\l .......... 511:1.'0
A 5/I;<Jil Rw;.,eJI ct ,\1 .. )Jlii7.'i
< A I;I'J'll< Ru"<tl .. 'it;Jl/
HlRJ.J(fN PA!'I.ON I' DOCUME]'; l'S
WT) J(/19<;8
OTHI:.R PUBUCA:l'!O!'IS
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ics of Nnrth America Equine Practk:e, vnl 2, No. 2, pp
439-459 (Aug. l 986).
Brewer, Barbara W., "Multifocal Neurologic Disca>c in a
llorse"; Equine Veterinary Science, vol. 8, No. 4, pp
302-304 (Apr. 1988).
Kobluk, cl al., (cds.) The Horse Diseases & ClicJcal Man-
agement, W.B. Saunders Company, pp. 459-462 aud pp
729-TJO (1995).
Schoondcrm3rk, ct al., "In vitro etl'ects of sulfadinine and
its m<:tabolite:;; alone and in comhmation with pyrmethamme
on 1oxoplcmna gondii"; Chemical Abstracts, vol. 122, p
570, 122:235089(n) (Mar., 1995).
Fenger, Clara K., "Update on the Diagnos1s and Treacmcnt
of Equine Protozoal Myeloencephalits (EPM)," Proceedings
of the 13'1> Annual Veterinary Medical forJm, American
College of Veterinary Internal Medicine, pp. 597-599 (May,
1995).
Amln;ws, Frank M. et al., "Differentiatmg, Neurologic Dts-
cases in the Horse Using Albumm Quolkut and JgG
Dctt:rmination," Proccedingh of 13'
1
' Annual Vt:knnary
Mcdic<1l Forum, American College of Vclcriuary Internal
Mcdiciuc, (May, 1995).
Podzamczer, el al., 'Twice-weekly tlier:.py
wrth to prevent recurrc:Jt
toxoplasmic encephalitis in patients with AIDS"; 1-Pharmd-
cology, vol. 23, p. 37, 123-102131 (u) (Aug., 1995).
Clara K. Fenger, Applicalior. of the Sruall Subunit R!bt\<,O-
mal RNA Gene Sequence of Sarcocystis Neurona toward
Ddineation of a Definitive Host and Serologic Studies
(P:J.D. dissertation, The Graduate Schoo;, University o
Kentucky, Lexington (Mar, 1996)) (believed 1o be unpub-
lished).
(List continued on next page )
/:.muum:r
(71) 1\llumev. Ag<'nl, ur !-inn Fuky & l.ardllcr
(57) AHSTRACT
I 'he present invenlton relates to ;r;u
for treating such dS horses, a!Dictcd with equine
protozoal myelocncephali:is or EPM The tbetapeuuc com-
,ompnse a eornb.nation of pyrimethamine anJ a
sulfonamide, preferably, sulfadiazine, in lhe ahsence o:
known therape"Jlic amounts of tnmc:hoprim.
15 Claims, Nu Druwings-
',
US 6,448,252 Bl
Page
OTHEH PUBLICATIONS
Moore, Bonnie ... Informal Survey of Equine Protozoal
Mycloenccphalitis," Kansas Slllle Univers1ty, Mar., 19%.
Bertone, Joseph J, 'Update on Equine Proto:wal Myeloen-
cephalitis," fDA Vet., vol. XI, No. III, May/Juu. (May/JlJtL
1996).
Fenger, et al., 'Epizootic ol' Equine Protozoal Mye!oen-
on a rarm", Journal of the Vctcnmtry
Medical vol 210, No. 7, pp. 923-927 (Apr.
!997) .
.. Tn;atmcnt of I.:.quine Protozoul Myelocncephttlitb" by B.tr-
barn Brewer Welsch, MA. DVM. ct a!. The Compendiwn,
North Americ-an Edition, Equine, 1599-1602 (1991).
''Equine Protozoan Encephhlomyelitis" by Jill Beech,
V.M.D., Veterinary Medicine/Small Animal Clinician, Dec.
1'.174, 1562-1566.
.. Comparative cm:<.ts of cotrimoxazole (trimethoprim -sttl-
phametboxaz.ole), pyrimcthamint'- sulphadiat.ine and l>pira-
mycin dunng rlviurlent infection with '/o.wp/asma gondti
(Beverly strain) in mice" hy B T. et al., /fl: .1.
Pharmac. ( 1983), 79, 923-928.
3615/:nodictyon, Merck Index, (1989) p. 530.
79'.17 l'yrimelilll!llille, Tlw Mcr<.:k (19lN) p. 1270.
HH74 Sulj(uliuzine, Ml..'rck lmlt:X, (1989) p. 1404.
Vl>l. (>'J, 196i>, p. 4749, Sec
'i0900p
Cite!IIIL'fll vol. 85, 1976, pp. t\6 47, S\Jc. 72303d.
1 -1'/wmwccxlytwmln, vol. 7H, 1973, p. 31, Sec. 527mh.
1 1'/wrnwcod)mtmics, vol. 77, 1972, p. 14, Set.:. 10933%.
1--Phurmacvdymunic.\, vol. 92, 19ll0, p. 63, Se<.:. 92:1558lp.
Chemu:a/ AhMrCH:t;, vol. 117, 1977, p. 60, Sec. 87:95742b
vol. 101, 19!14, p. 352,Sc<.:. 101 HTB!JI.
Chemicttl AbMracts, vol. 99, 1983, p. 20, Sl:c. !JlJ: l33330v.
63 Pharmaceuticals, vol. 96, 1982, p 373, Sec. 96 408451.
1-PharmamhJf{Y, vol. 123, 1995, p. '>o7, Sec. 123:Jfi211Ju.
Chemical vol. 123, 1'l'J5, p. 38, Sec. t:B:71277<l.
C/lt'/1/ica/ vol. I 22, I <J95, p 44, Set'
122:2'>o5089n.
Clwmil'lll Ah.\lract.,, vol 122, 1995, p. 44, Sec. 122:45806w.
l l'harmacology, vnl. !20, 1994, p. 33, Se<.: 120:45263a.
1-l'harmaco/()g)', vol. IH!, 1993, p. 29, S.:c. ll8:182836n.
"l'harmacokinctic:s oJ' intrnvcnottsly und orally
nrim<>thnmme in horses" by Cyril R. Clarke, Ph.D.
ct al., Am. J Vet. Res., vol. 53, No .. 12, Dec 1992.
"l'harmacokin..:tit-s, penetration into ccrcbrospim11 l1uid, and
hundtologic effect> nftcr multiple oral administrations or
to horses", by Cyril R. Clarke, BVSc, Ph.D
ct a!., Am .1. V.t. Rc>s., vol 53, No. I 2, Dec. 1992.
"In Vitro [fil:cts of Su1t'<ldiuine and Alone
and m Combindtion With l'yrimctharnlllc on Ji>.wp/a.>ma
f:OIIdit" by De Vcn cl al ,l\11/t
mit ro/?ial Ag<'nl.> and Chemolli<!mpy, M,lr. I <)l)'i, vol. w,
No .. \ pp 763-7t>5
New to Study thc Ltli:tt ol
on lrJxophl\lllO gorulii. ol' Sull'at'uxinc
and Su li'athumc lndivicbally and iu with
nnd Study ol' Ckid:unytin, MctJt>nJduo!c,
and n" by D.G Mnck t;;t a!., Anlilli/t:rohiai
and Ch<mwthertlpy, Jul. 1984, vol. 20, N<>. I, pp
.?6-3(1
l'I'M Semmar, The J-lnr><', Nov. 1 '1')5, pp.
l'roto/.u,J! Myd1t1'> \Vork.'>hop. Sunlllldf)' r'" the
Ilorscrnau" (/ra_nrm !ockev Clul> Re.search i"oundalton
Seminar, Mur. I <;J6
David S. cl uL, "Evaluation o: AntJ-..:o..:cidial
Drug;,' Inhibition of Neo5pora cartinum Development in
Cell Cultures," The Journal of Parasitology, Dec. 1989, pp.
990-992.
David S. Lindsdy a!., .. Demonstration of Synergistic
E!lccts of Sulfonamides Jnd Dihydrofolalc Reductase/
lnh!bJtor;,/\gd;n;,tNe()spora crminum
Tachyzoilcs in Cullurt>d Cells, and of
Mutants Re;,istant In AJVR, vol. 57, Jan.
19910, pp .. (Jil--72 .
J P J)uh..:y eta! , Antkocc:idi<tl Actlvlly ol'? -i'>u!f:trnnyl 4,
4-DiaminodiphcnyhuJf,mc. Sulladu.inc, l'ynrncthammc
and Clindarnycin in Cats lllfe,ted Wtlh J(>:rc>pla\11111 K<mdh,"
The Canr1tlian Veterinary Juurnal, vol I H, No. 3, Mar. 1977,
pp.
Uanicl l'udJ.amco.:r .:t a;., ... 1\vtce-weekly, Mamtcnancc
Thcrapy with Sulfadiaziuc l'yrimctharninc to Prevent
ToxuplasmJl: in Patients with AJDS,"
Ann.tls of btcrnal Medicine, vol. 123, No 3, pp. 175-180
(Aug. 1991).
B.T. Nguy\:n ct al., "Comparative of Cotrimoxazole
xtuok ),
l'ylirndbamtnc sulpbndtuinc .md Spii<Jrnycm Dunng
Aviru'ent lnll:ctioll w!lh '/mop/a.\111<1 gollllii ( Bncrly Strain)
UI Mi..:c," llr J l'h;um.tc', vul. 71J, l 'JH:\, pp. 91:1-928.
(';;rolll:irri, cl at, "In V:trll ul' AntllnJt:robJH\
Ag..:ntl> Agamst "Jv.wpla1nta wmdtt," rhe .lc>IJrnal ,,f lnke-
tiouo vo!. 157, 1. .Jan. l'IH8, pp. 14- 22.
llarlcy G. ct al., "Effect of Pyrimethamine and
on the f-ine Structure and Multiplication of
Toxopla5nw I{Ondit m Cell Cultures," The Jouwal of Para-
sitology, vol til, No 4, Aug. J97'i, pp. 704-712
I.G. Mayhew, "Latge At:imal Neurology: A Handbook for
Veterinary Clinicians," Lee & Fcbigcr, Philadelphia, 1989,
pp.
Bru<:.: G. Ccllin ct a], .. CucddJan in AlDS
,!I'd "I he
1\kdtt:.tl C'lti:il:;,ol :--lt.:lhAtncr\.a,vol 7(,, t,,;, !,Jan. l'l'/2,
pp :or, Y\4.
( 'l<ra K. Fcngc r l'l a: , ''Idutldic d\.un tl Opo-...,um:-. (J)tdef-
Jilll.\ l'ti'Ril>llll?a) .,.., 1 he Pu:atiw Udinitivc I-lo'>l of .\arco-
' pti., fl<!urom,," Jour71al of Par,tsitnlogy, vol. HI. No 6, Dec.
1995, pp I) 1 (J-'J I')
cted by examiner
' .
. '
.
US 6,448,252 B 1
1
TRJ<:ATMENT OF EQliNE I'IWTOZO/\L
MYEJ,OENCEI'HAIJTIS
2
to eat) and may del if it is ..:hoked with feed
<.:oming from ami biol:ncmil-:<1 blood
arc usually in the normal range.
This a .s a continuntion of U S ;tpplicntion Scr. Nu
OQ/06C),CJ56 flied Apr. 10, I C)QR now U S Pal. No. 6,255,308, s
in..:orpomlcd by rdcrcncc til t's enti1ely. wh:ch is ''
continuation of US applicdtion Scr. No 08/61C\5071lle<i .lui
of EPM IS based on dinical sigrLs and on testing
of the cerebrospinal fl:nd (CSF). Originally, the
diagnosi> wa.-. ba>cd on the presence of antibodies to Sar-
!leurona in serum, though it is now known that a
pos.ilive tesll'annol he used to make a diagnosts; such
pt''itltvc simply illl.licntt:s t:llpusure to the parastte,
17, !996, now US. Pat No 5,7\7,L76
J. FIELD OF THL INVENTION
prcs(;nt UJvc.:utiot! to .:ompostttons and meth-
ods for tn:allug such affiicted with
cquim:, pwto.wal of EPM. EPM is a
debilitation neurologic disease of which cau alfc.:l
the brain, the brain stem, or uny wmbinatiun t1f
these three areas uf the equioo 's t:entrul system
EP:vt is caust:d by inl't:ction hy the para.,itc
neurona (recently referred tn us Sarcot'_)'\li\
fa/cacula). There no vaccine or approved animal dn:g
product available for effectivdy treating 11is disease .n
horses.
2. BACKGROUND OF I'JlE INVENTION
Nthough the ,wcl cftccts of lii>M havu lJCun
recognized the 1970's, 1\ not until 1991 lhnlthc
protozoan parasite that EPM was cultured from a
horse and given the name Sarcocy5cis neunmu. 'l11e horse
an aberrant, dead-end host, as tnfcctious forms of the
p arasitc arc not passed fwm !Jobe tu hoH;.:: or ffl1m
horse to a definillvc or true intcrmcdhtlt:
investigations indicate that the uf the Uptls.-;urn (the
dchnitivc arc source of the l\lr horses.
EP:vl lfl much or North 1\lllt;lil.!d. Mtrwys
co11dul.!lcd in t:tmtra 1 KenttH. ky nne;; in
ami the nl Ohio and Oregon, have tcveakd tlmt
appnlXlmatcly fifty percent (50'fr') or the hur!IC!i in
surveyed aretts h1tvc been exposed to ih(l lthuw-m>tetl pru
toman A po'>tive serum test iPdil.:ntes cxp<lsUre to
lhc parasite, not the prcence tlf an active form
nf the The incidem:e or the llCll"l: dis<:U!IC is much
lower.
[n studies that looked at the distribution uf
cnses it found that climatic liu:tors mny
affect exposure rates; i e., an i:Jcrease of frecrlng days ur a
very hot environment was associa:ed with a in the
numbers of horses exposed to the para<;ite EI'M tn
have a distribution, although outbreaks have
occurred on farms in K.:ntucxy, Ohio, lndi41lll, M khi,I\JH
I'lorida.
A horse;; of any age, breed, or sex :nay be alkctcd hy I cP\<1
The disease bds m a horse or two month., of age.
Ju not necessarily preliUnt,.'t: ul' the diseuse. Cerebrospinal fluid
testing is now believcu to be tht: most useful to ass1st in
the diagnosis of dtwuse in a live horse.
Curreutly available lfetdmcnt of horses with EPM is
expcnsivc and typic31\y requires a duration or at least ninety
JS (90) days. ln some t.:ascs, trcallnent !1151!1 iudcliuitdy. l11i,;
'urrcnt nu.:ut involvl.l!llhe adnptutiutJ lit' tub lets
lor human l1nll\, tahletJ> arv atlrnini!<
along. with luh!t:th u trimcthuprim.
sulthn:tmid.: <Xlllloinatinn. 'l'ypit:afl}', lht: 1Wl1 types nf t,thlcts
in arc und plncccl rn susp.:ns1or: tor oral administration.
These mcdic:,tinns should administered one hour pnor to
lced:ng hay and are accompanied with frequent, periodic,
veterio11ry, neJJrologic examinations during the treatment
pcriou.
:.:t l)isconlinuation uf therapy h, bawd on adnJin-
mtrntiou nf medication thirty days beyond the plateau of
..:linical improvement or disappearc.ncc of antibody to the
prtlHllwa fmm the CSF. Suboptimal dosing or intermittent
_U) tbcrupy hill> no pn.wen efficacy.
Adver!iiJ of therapy may in dude anemia, abortion,
diarrhea tlmllnw white blood Both medication:-.
lur trcatrnt:nt uf bPM :ntnbit fohc a<.:ttl metabolism. Unlike
horses, hHWI/Vt:f, the protozoan is unable to utilize pre-
.!> udtl. Supplcmcnt,ltion with fnh.: <ll'Jd or fohni<.:
a.:uJ ( Wlllg orally, once d:1y) anll!or brewer., rnay
help side It 1s
tbal folic actd not he administered n1 the same time as the
pyrimethamine bcc,n .. se of competitive inhibition and
l'J :th"()rption.
Tlw and sporontocidal eJI.:cts of 2 g
s11lfudiazinc with 50 mg pyrimethamine in a chloroquine-
rcsistnut strain of Plu.;madium falciparum JS disclosed in
('htJmical Vulumt: 69: 50900p (1968). Primaquine
IJ' tliphn,phatt:, pyrirncth.wlmtl aud sulladiazinc were said to
causal prophylacti<.: al:livity against rodent malaria,
Plownodium hr.'rj?hei yoe/ii, as d!;,closed in Chemical
Ah'ltracts, 77 109339h ( 1 CJ72) A tbree cornpoocnl
cnrnpusitinn nl pyr1rncthurninc, suira,lluinc ami
,, !J('l for lrcaling rodent is in
Chcmi..::il Ab..,lraels, Vnl!Jmt' %: 4084 'it ( 1 982) Similarly,
<;odium h"' been ll> cnh<nrcc the
of ccrtnm anltint'e(..:live drugs infection' cnu,ed !1y
as well as one in 1ts thirties. In fact, any uorse demonstrating
neurologic should be col!side!ed a candtdatc s;.
for EPM affliction.
pyiinJctharninc-sus.:eptible or pyrirnelharn inc-re;,1stant
strains of P {alcipamm and P vrvax in owl monkeys Sec,
Abstracts, 'vOlume 92: 1558lp (1980).
There &rea number of articles de;.crihing the treatment ot
Toxoplasma gomlii with pyrimethamine and sulfadiazine,
hut Bl r:l'in thai uses n very lurge umount of sulfndinine
relative to pyrimethamine. See, e.g., Chemical Acstracls,
V0.umc 78: 5270Ss (1 97 3)(1 mg/kg pyrimethamine and 100
mg/kg st.lfacliazint for m1ce), Chernic:d Ahstracts, Volume
85: 7=:303cl (1 076) (2 m!'f.-<g p:ynmethamine and toO mglkg
;,dfadiuine ltll cal.;,), Chcnu.:al Ab;,iracl,, Volume 99.
Climcal signs of the condttJOn depend on the location of
the organism within the central ne:vous These stgus
incluc.e weakness, malpos1tion of d limb. muscle atrophy,
ataxia, or ''wobbling'' a;,d/or bead till with .tsymmctry otJ
oi' the lo<:c (q;, eyelid, e.tr, 11r hp). /\seve: ely
!10rsc m.ty hc.;,,me recumbent :md unab;c to
not traceable to orthupechr: di;,easc '" .:oml1!n,t!ion ol lt:c
sig11s may :>cr:vr wit:t U'M Other :lliU>,Jd rnay
alStl OCl'UL '' L\Y\.10v (I%<), 1\hlnl>"-<, IUl
(''ltd),'\ I AI>., II n,'l'-'. Vulurn<: : 2? ( 1995) (I
ng:i..s .wd "lJ mg.kg '<.lind:,uiPc lor
cu"t .... , <In aP.'ccted nor"!.' ltni..! (llut wttl! d
nor,c1al ppctitc, ah:lnugb 1. ra: :,, c:y-.,'h"grc ( .. c , Ul!:.l>lc
'
. '
us 6,448,252 I31
3 4
particular, prior <.:omposition; for the treatm,nt of EPM
1nvolve three-component mixture,, tncluding
pyrimethamine, sulfadia1ine and trimethoprim. Moreovtr,
where prior compo,itions contained pyrimethamine and
Chemical Abstract>, Volume 122: 253089n
(1995) Chemical Abstracts, Yol1.me 123: l02Dlu (!995)
describes the daily of 25 pyrimethamine
a total of 2 g sulfadial.inc to prevent toxoplasmic
encephalitis m AlDS patients.
The patent literature indu1.ks many descriptions of meth
o1.b treating protoz.oan-med1ated diseases. McHardy, In
5 su:fadiazine as the active ingredients, such compositions
very small ratios of pyrimetha:nine to sulfadiazine
limiting their ell'ectiveness to treating malaria only and
hampering their usefulness in other pathological conditions,
like protozoan-mediated diseases, especially EPM. The fact
U S. Pat. No 5,273,970, sta:es that a proto1.oal disease,
toxoplasmosis, may be controlled to a .:crtain extent using
together with a sulfonamide. This patent
asse:ls that baquilopri:n cau be for the treatment and/or
pwphylaxis of protozoal infections in animals, including
Although tile baquilopnm can be used as the sole
active ingredient, ll can he with a
midt. A long list or 'ullaillc >ulforJarmtlc,o., IS prOY!dell, 15
prcrcrah!y ,uJfadia'l.:ne,
,ulfudirnethoxint:, ,,ulfamnxolc, ur 'uifallirrll-

10 is that there is currently no approved drug or drug combi-
nation for !be treatment of EPM.
In U.S Pni. l'<o 4,599,416, granted to Kompis, a process
20
for the prepardlion ol' aqueous
and pot"nllHlOf' for lht! treHimenl or IJacterial
ir:fcctinn' in humans< nd ammab ,, dcs.:nbcd A long lil of
poleuli<ll ,uiJ\>n,,mJdcs j<, provtded. Tbe urc
dcs.:nbcd ,,, tknoting compound, thai iucreuse the antibuc-
25
ltri<Il of sulfonnrnidcs more than additivcly. An
equally Jong-"laundry" or su<.:b potentiators is provided,
wh1cb iududcs tnmethoprim and pyrimethamine Other
ptenl& dealing with proto:.wnn parasites include: U.S,
No 4,293,54 7, granted to Lewis ct al. for the treatment of
malaria; U S. Pat. No. 4,340,609. granted to Chou
30
protowal infc,tation.-.), U.S. l'at. No. 4,368,193.
Argoudclis cl al. (malaria); U.S. PHI No. 4,728,641, grunted
to Tubaro al (proto1.oai inkctioos gt:neral\y); U.S. P<Jl.
No. 4,992,444, grnntcd to Steven' 1.-l al. (trypanosomcs and
malaria); and U.S. Pat. No. 5,486,535, granted to Marr et a!.

(Toxoplasma gondti).
Beech, in Veterinary Medicine/Small Animal Clinician,
pp 1562-1566 (December 1974) described a condition in
cighl (8) horse> wllh signs t>f m;urologkal disorder. On the

tt:at toxoplasmosis was involved, the author
suggested that pyrimetbarm:1e aad sulfadiazine, used suc-
against toxoplama in man, might be w;cful m
horse,,
3. SUMMARY OF TilE INYE;\ITION
Quite it has now heen discovered that an
ell'ect1ve, convenient method of treating EPM is realized by
the to nn equine suspcetcd of hcing affiictcd
with l:.PM of' thcrupeutk amouoh ,,f d 1tl a
preferably Inc rdativc weight
ratio of pyrimethamine to the mny range from
:1bout 1:!0 to nbout 1:30, preferably, alxml 115 to about
l :25, and most preferably aboul 1 20 in lbc CdSC of a
composition comprising pyrimethamine and sulfadia:.dne.
11 should be pointed out lhat :he compositions of the
present invention do cot contain significant amounts of
tri:ncthoprim, certainly less than about two-thirds of the
weight arnouut o! sulfadiazine ptesc:Jt. Preferably, the thera-
peutic compositions used for tbe treatment of EPM arc
free or trimetboprim, most preferably having
no trimethoprim al all. Similarly, the methods of the present
invention do not rely on the of signitlcanl amount;,
or trimcthoprim in succe;,sful treatment of EPM,
u'ing the pyrimethamine and n sulfl>nalTilclt: ai>
lhe prmcipa; H<..livc lllgrcdicnts pai:Jo!ogic
namely, the organi;,m Sarcon'.\tis neurona in EPM. Hence,
the methods of the present nwenlion do nul mcludc the
co-administration of known therapeutic amounts of tflme-
tboprim.
In a preferred embodiment of the invention, the affiicted
equine, e.g., a horse, is given a daily dose of pyrimethamine,
which is equivalent to about 0.8-1.2 rng per kg of equine,
most preferably about 1.0 mg per kg. 1be subject 1s abo
given, concum:nlly for the convenience, an amount
day of d whkh IS equivalt!nllo aboutl5-30
Wchch, B B recommended tht use ,,r pyrimethamine
(0.5 mg/kg), <:ornuwcd witb a 20 two-part rr.ixturt' of
( 16.7 mg/kg) ,UJd trimcthoprir:l (3.3 rr.g/kg), In
horsts lPM Wcl,<::J, !l. B in 1 he
Compendwm North Amei'lcan Fdlfion, l:'iJlUtle, Morr,,, D
45 rng per kg of equine, rnosl pre!crably ahout 20 mg per kg.
D. (Ed) (1991) pp. !599-1602
'lwo nrncles by ct al, whch !tppcan:d m American
Juumal of Velermary Research, Yohun.: 53, Number 12,
pages 2292--2295 and 2296-2299 (December 1 992),
the pharmacokir.eti<.:s of :ntravcnously a:1d or,llly ndrn,nis-
pyrimctbammc in horses. The first ar!idc, al page
2292, o,tateo, that c:mic,,l reports ir.dtcatc the poss1hle value
of trcalrncnl of horse> with prolozcal encephalomyelitis with
pyrimethamine in <:omhinanon with and 'ui-
f<r.amidcs On "land, tt:c second artkk, at
2::'
1
)1), C<>m:Jucic-, lhil the ora] of ]
pynmcthITI!llC/kg cncc " J.1y lnr J(l dayo; ,;""'
col ;>rcs.:nl a toxicological prohlem .o hor"''"
llcn,c, despite a great dc:ll ut ptl'>l n:l(! on-going dlnrt,
thc:c re:m:ins an JPlfulfilkd need for <l lre;otrncn: for U'M-
affiictl:d cq:Jmc>, ?articularly horses, whi<:b b not onl\
etfcctivc but i.s a:so cunvcn:cnl to to rnaximtzc
curnphltlCC and rcuuc<: the emergence of rcsislanl st1ain; In
OtK'c daily :tdrnirnstration of the nctive
evury fiiiWiinf!, on an empty for at least about 3
month!., preferably about lO IHO days, j-; <;;1:lkknt to lrl'al
the In <,O'l1C how<:vcr,the treatment regimen
50
ca 1 lao;! inddimtcly, l1>r tl:c rcrnai011h life of the
h>r of the therapeutiC Ct>f:lJlOSI-
lion may be given orally (thai is, hy mouth).
II should he apparent lhal .111 object nl' the plc,enl 'nv<:n-
tion is the lrcalrncnl or equine prolozod: mydoencephalilis
5' or EPM by providing a veterinary composition comprising
pyrimethamine and a sulfonamide, provided that the com-
position 1!ocs not also includ.: signilicant awount' of trime-
thoprir:l. By "not also .nclude .,ignificant arr:ounts of tnme-
thoprim" mer.ns thai acy tr.methoprim present in lhe
<><l vcil::rinnry compn<>:li<'n shm!ld nnt rc:rch any xnnwn thcrn-
peulh' leve.s of not renchine an
.<rnount by wcigh:. wlncb l'-i equJv,Jlect to ahi-.It two-th.tds
the d the ,<<.:!ive su!fonnrnide, prcferah;y le<.<, rh:m
a::.out one-bnlt .11d P1ore prele,ably :css than ahout,,ne-thml
'-' of the 'ullondm1de. tb .. vctcrinctry compo-
'illion of the prc,cnt lnv.:nt:on (cr :ncthml <::
tr<.atmcut of EPM) free o: tnrncthopr11n.
US 6/\48,252 B 1
5
Couvcnicnl dosage formulations of the present invention
are also cor.templated, includmg solid and liquid forms, and
umt dosage forms comprising about 0.3-0.7 gm
pyrimclhaminc and about 6-14 gm sulfonamide, preferably
about 0.5 gm and about 10 grn sulfonamide.
These .md other ol the invention will bcmme
appawnl to !hose or ordinary >Kill in the arl, especially after
constderation of the following dctmlcd dcscnption of lhe
preferred embodmlents.
4. [)J..rli\JLED DESCRWJ'JON Of' TilE
PREI'I::RRED EMBODIMENTS
The instant invention iuvolvcs, in a prefe:red
o;;mbooimenl, the administration or tin oral cun-
taining ami a sulfonamide,
design<:d to ovcn.:ome the 'horh.:onung' ol
currently available treatments of EPM and to provide a norc
cllective drug combination for ami other
mfcctcd with an organism of the genus 1\s
prevmusly mcntJt>ned, pyrimethamine may he gJVcn in a
preferred dose of about 1 mg/kg equine with a sulfonamide
in a dose of about 15 to 30 mg!kg equine, preferably 20
mg./kg.
6
b a .'>irmbr manner, nuf!lerm;s sulfonamide1. mav be
suitably utilized in the pres<ont invention, indudmg those
previously disclosed in U S Pat. No. 5,273,970, whose
diSclosure is incorporated in its entirety by rcterence herem.
lo particular, those sul!<Jll<lmides having a dinitrogen aro-
matic ring are espec1ally useful. such as acetyl
If a met boxy pyr a zi ne, N- 2- form ylsulfisomidine,
sa I a zosu I fad i m i eli ne, su Hac h lorpy rid a zine,
sulfatlimet!Juxwe, sultadoxine, su lfalcnc, sulfamerazine,
n sulfameln,
l>ulfamethoxypymlazine, sulfaper,u<o, "ulfaphenawle. sul-
fapyrazine and sulfisom,dioe.
!:->till known to bt: u<;efuj in veterinary
can b.; indudiug
'-' phthalyl,ullac:clarnlllc,
att11" 1<>lc, 'u l:'abtnt.<tnJJ(k, ace! am ide,

sulfanil.t m1dc, sulfanilarni-
domethantsulfonic acid triethanolamine salt, sulfanitran,
2u sulfapyridine, sulfathiazole a:1d sulfisoxazole. Tre above-
mentioned sulfonamides may be used in place of the pre-
tened sulfadiazine or m addition thereto.
11 hdfi been [ounllthal a 30 rnL dose;: o[ an ural
25
(.,uch itS thai tlcS<.:nbed, below) daily on an empty sloma"h
will provide adcquatc thNng lln the lrtatmcnt ol' EPM thai
neither pyrimethamine nor can tree! <!lone.
Sine<: EPM i1-> a pmlotoal inrc<.:llon ol' the nervous
the cln1g combination to ,
0
the CNS and treat the proto/.llal int'cctiuu
The present compositions may be administered by routes
well known to those skilled in the veterinary and formula-
tion Therefore, nlrhough the pyrimethamine and
sulf<H.!ittzme, for example, a1 e conveniently administered
orally, depend in)!. u:1 the circum;tan<'es, the pharmaceutical
compo;,ition m;;y be Wmimstcrcd parentally, topically,
;ntramuco>aily (e.g, intnlv.tgina!ly), or by other routes
to ,k,:lcd m tin., art.
Cornpo.,llitln<; suitable lnr ,,rnl adntimslralion, in uddition
to indud...,; gt:h,
hoi uses, or prcpnra:ions u1 the tonn of powders, granules, or
.<; pellets Preferred orally administered compo&itions ir;dude
suspensions and tablets
In g,enural, the to be adrlllui,tcrcd llirlY 1-<>lll-
pnse;: about W-20 gm pyriruel!HlnllOl' ami about J.'iO ()()()
gm sulfonamide;:, prefcwbly 200-400 gm per
hler of I!Omposihon. The;: liqu1d or cmnptiSition may he
prepared in unit dosage form depending upon the minimum
;.ize of the <Jquine. Such unit dosage forms comprise a
rebtive weight ratio of pyrimethamine to sulfonamide in a
range of annul 1 :L'i to about 1::10, preferably allout l :20.
'1Ypically, the unit dosage forms contain about 0.3-0.7 gm of

pyrimethamine and about !i---14 gm of sulfonamide, mosl
0.5 gm pyrimethamine and uboul 10 gm
mide
1\llernativdy, tl;e eomposilmn may be ad:ninistered
parenterally by sub-cutaneous, intramuscular,
intraperitoneal, or intravenous injection, or by implantation.
The can be given as an intra mammary injection
whereby a suspcrl'.;inn or is introduced into the
udder.
l'fJarJTJaccutt..:ally camo;;r.; in the com-
:ll">ilion" ol' the tnvcniJ<>n arc rnatcril1b nKom- prc,<.:ut .uwntiun h.ti> been lound to 'uccc;..,:ully
inhibit lhc growlh of th<: organti>tn Srm:oq,fi., neumrw i11
u;. mule;., fl<Hilc' :md ll ha' been
ohF.crvccl that the preferred sullonnmirie. hn'
rcs:Jited in !">eller than about a 70% r11tc of ct;i<.:acy In tact,
'Jtcl:dcd lor the ol tldrr-in"lt'fHI)!. the mcthl'<HTICfll
my h\- liytml, '-otl,Jd. 01 whilh arc
l:therwi,,c inert or rnctli..:ally .c..:cplahk >111d ate compat1hlc
With the liCtive mgwll;cnl'> The >;arne !ipplie' for .wy added
the effectiveness of the present <.:ornpnsillons and methmls
appears to be at least abmll 80% or 90% of the and sc
probably eveo higher.
exctp;enb
For nral ad;ninislratHYJ, tine powders or granult:s will
eon\i1m dtlut1ng agmts, for example, cakium <.:arbonatc,
eakiurn phosphate, mineral earners, etc., dlbbursmg ami/or
surl"aee active tor example, poly&oriJJtcs, and may be
prese;:me;;d in a dre;;nch, in water or in a syrup, in a bolus,
ln line with the foregoing, ll IS within the contemplation
o: the present invent:on to employ \-1lmpositiom. uliliLing
one or more sulfonamides andiur pyrimidme dcnvative in
treating EPM. Examples of other pyrim1dine cteriva-
but "''t lmllit:d to, 2,4-diarnino-'i-
hu:ly'pynmtdim'> in I he p!1cnyl ri:w,. 1 s
S ("\,'i dinwthoxy-4-rwiiHxycthuxy!,cnt.v)-
pvnmid!;rl'(lc\rnxtprim) Hlld :,.1 dr,lrllii'O-'l-( '\ -;,
d 1t11C IIJoxy II h il'hcn/ yl)-pyr. tn Hllllc(I'IC iop in:)
S.tll tth..:r "'ci\JI rnay h<: 2,1-dlarninn-'-(4-
h;or.m-1,5-dirnethoxyhcnzyl)-pyrimidim:, 2,1 dt mino-'i-
[:l,.'i-diatboxv-4-(pyrrol-1-yl)-hcnly.]-pymntdinc, 2,1-
th.< mwo- 5 -(3, S-cli rmt hoxy-4-d i mt thy I ami oohent.y I)-
p y r nr: 1d m i no -5 -( 3.-t-d imc:boxyh;n 1.y 1)-
pydmtdinc (C:iuvcridiuc) and 2,4-diaminv-5-(2-:ncthyl-4,5-
dimcthoxyb.;n,_yl)-pyrimidinc.
I' paste, or in capsules or sachets in the dry slate or in a
or 111 .1 suspe;;nswn in water or
Whcre desirable or
'uspcnchng, tlnrkenhg or emulsifymg agent.< Clln he
1r< h1dcd It' irllcndcd fnr ornl L'>C, a will he provided
'' wnt; rclclll.uc mcm., tc rcg:trgtanon. [",,r CX<lfllpk,
11 be w.:.ghe<i v.i1h ; hl':1vy rlcn<ty 'uctJ as
in>1 or or the ur :1111y oc by Jh sha,Jc,
l'ot l'XHmple, hy whd1 ''" ,g :dkr tdndni,tratinr.
nwy co!ll:lJn ;tgc.nl;-, H'l ml:llll'
,,\ or ca:cturr, sodi1;rn mc.hylt'c'liulosc:,,
hyclroxypropy lmcthylccllu!;'"" b<t.>cJ vegetable gum5,
sudium or stJr(.b g:lyc:olatcs; gra:1:1lating or
US 6,44R,252 U 1
7
binding such "larch in the form of ;nUt'ildgc,
derivatives, such as methykellulose, calcium stearate, talc,
gelatin or polyvinylpyrrolidone; lubricating
SUI.'h as magnesium stearate or steanc acid.
Other which may be.. included for
exJrnple, rnedk:llly inert i:lgrcdieots, e.g solid and liyu1d
such as stMch or ca.tium phosphate for tablet>,
or capsules; olive ml or ethyl olea!<: t(lf -;oft cr!p-
suk'; W<ltcr t>r vcgddll:c ud fo1 \u,pcm.Jon' or crnul-
.... luhrkdling ,lgt.ul:-. <.;uch as tall' or 10
gelling agent" such coll<>idal clay'; tbtdwning agcnh
a' gum tragacanth or 'odium :lginatc, tkdu,ung agcnb
such"' liyuid parallin, fixLd oib :1nd 'urldctJnh <!lld other
lhcrap"utically a<:clpl<iblc ingrnlicnh. as
prcbcrvativcs, hul'lcr'>, and ,Jnti-Oi<H.l<tnb. wluclt
drc useful as ""rriers tn formuld!ions. When
other and/or nutrients, ;uch 01
likc, unlcs; conlraindic,ltcd, may also be
It is also to be understood that while the preferred
formul<Jiion is administered once a day, it r:Jay be gtvcn two 20
or more t:rnes a day. dependmg <Jn the circumstances. lt
be undcr;tood thai while it is coownknt to admil:-
islcr the pyrimidine and sulfonamide concurren:ly, they can
given separately with cyual dlicacy However. the other
optimum such the adnwu,tr,ttion ol' the t:rug co
on <1n L'IT!ply, prt'!cr,!lliV .tl ka.,l ,Jhoul nne hur
the J'cd shlluld
A lurthcr ol the mvcnlion may he
!'rom the lnllowlllg 'pcc>llt: l'hc'c 'JlcCtll..:
<He provided lor il:ustrauon "nly lind ate nnt to -'
0
regarded as restrictmg the invention in an} W<Jy.
5. EXAMPLES
Veterinary compositions enectwe for the general treat-
ment of El'M arc provided, below, in the form of an oral 35
suspension. The amounl' of component are hased on a
liquid su:-.pension having a total voiUI'!e or about 1 liter AI>
mentioned above, a useful dosage, e g., lor a 1,000 p:mnd
inl'ecwd with .'wrcocy.,/1.\ nf!urtma (a1. evtdenced by
the prci>Cnt.:C of I he pruiOtOJll t,l a sample frnm the subject's 40
ccrdrospinal lhtid) j., ahtHII 10 mi. ol' ,tJc 'l"pcnsion, nnl'c
.1 day, on a11 empty .-.. tomach m the
L llSF
?, Sulff,(}o!u'lr.c Sndmt1',
J PSP
4. Soclllm NF
'), Xfnth!.J.l gu''l, "\t'
6, H'W. Ni
7. '\;'l
'( Ycll""l v
tlll\ll
'J H 11,'\(> "'!,
1 . .'tlh'> ,, ll',' ('{f '\I
j, 1
1
, d t' !, , -.,p
l'crl.it<1
0 r S
f-;7!!,
10 D'/ g
'2 22 5
l.l
lJ
:. ?8
"':5 5t. ll'l
t:Jl,
,ul II'
:. ,;,,
,,
l'rclcrd>ly, the dnc'> uu1 c:onJ,Itn -;uh,t,lnltdl '"'
nmouni:, ofnrtlllf,d 'ugar' Mr"l prdcrahl_v, the '"''TII"''JIHHJ
ss -;ubstanlially free of sug<tr' a-; sorJJr of ihc ,\!1-
fonamides may he sensitive to the ;Jreqenzc of rwi\Jrally
sugars. OptiOnally, t(Jiic ac:td may :ilso be adQio-
JSicrcd to the >UbJCC!, either concuncntly or al a separate 65
llmc Typ1cally, the suhj<:ct may receive about 40 rng uf [ohc
Jcid per 500- tu !000-pound equine.
s
A stepw1se procedure for the preparation of the orl
suspen-;ion is provided helow
Weigh out the powders and triturate from the smallest
quantity of powder to the largest quantity of powder, so
that all powders are evenly mixed together.
2. In a '>eparate container mix all thc liquid; and add half
the volume of wMer for one liter
3 Mix malcri,tls prepared in steps I and 2 in
,t !urge mnlaJJlcr and q.'>. to nne ],000 liter with .be

I llc<tt !he "'"pcn,i<>n on a mi<'r<>nizer for 1\vc
hci11g n:rtnin \:ltd i!> evenly mixed.
5. l'.td .. Jgc the l111:tl pruducl und rcl'rigcr:Jic wh<:n 1101 Ill
u,c. !'he be well prior to usc.
As further illustrations of thc compositwn of the
invent10n, the following descriptions of suitable alternative
formulations are provHled
1 4
}
t Ht Nl
4 Xl!nlhn
., '\t
j t U.a\''lontt,

'1. l'unlkd waltr, I!Sr
Per L1lct
,;,<,l.,l4 t
I -\.}4 i-
:: l-

J'}lllf.
.,,r, ml
:,t,l>i u.L
(.' .,, l<J J,-!10 !fll
The preceding formulation provides an easy to use paste.
Formulation C
llSP
! 1\'lintclb..>ttnitut, tSP
' S4:WiutH Htnt,totW. Sl
r. Xamhn 'II
'1. ""\UiM"*W4!d
t. t ,trmnc1 'H
."!. lfH, Nl
l l 'Sf
CO!Tlpuncnt
J SulfRdin7me Basc
1
USP
2. Pyritrctl'rt1PH1C, l):-,p
3 .Sod.um Bcn7oatc, 'JF
4, Nllt!CI'iWt!t!l, NU
L.;..:tose. N1
po ,, n,
I ""i l .. .. l L I,-,:, ' :"'>P
''bill {'i <til' I'- t .,p
\()d I II jft_ !''! (. ..... !
t l'H1:1l'Wt.d ;-..,
j,<.,(o)'L'. '\,:
l'et Liter
_133.34 g
1J.J4 g
f:\

II:
'."b .nl
:1(1: lll
tl ml
Per 1000 g
:<:'3:14 g
l'l , .. , g
2.22 g
:o 0 g
tdFC1 g
\I;F' ')t,
1 t: ., l
The prcccdwg two formulations provide powders, which
can he convement:y d1VJded mto ir.dividual each
contaming 30 g of the inventive composition.
.
. .
'
US 6,44g,252 B 1
9
(ompJnent
Fcrmulahor F
l. OSP
2 h'!W'lctul.fru;c, CSP
3. "\F
' >,,,nlhn g\ . ' sr
:-. !\"t,l l\'1
f, ('J>fPTtd ,lg, 1'\!
!Jd\"l<)fbjlt "-;('
9, Fnt.ftt:d W<llC'!, t St>
f:-riTH!hHWn (I
... Sul(udimelho)l.[nc, US>
2 Pynrnethnmint', USP
3 So<..1um Benzoote, !'1(
7
4. Xnnthnn gun:, !'. 'F
5 NF
7 caramel ftavoting, NF
8. Polysorbnle 8C, NF
9, Purified wnter, USP
Per Liter
333.34 g
13.34 g
2.:!2 g
5.,Jg.

!0 ml,
6 ()/ l'tL
.., ... to J,oc:J rnl
50G.,Q g
16.67 g
2.22 g
s.o 1\
10.0 g
5.56 mL
6.67 mL
q.s. to 1,000 mL
Only the preferred embodiment'> of the invention and but
a t'f ''" nre and described in
ihc prc-.,cnl dj,,,;l,,;;\Jrc. It :'lobe understood that the inveo-
titlll j, ol in variou'> other combinations nne!
cnvir:,nmcnl' and is enpnhlc nr changes and
within !he ...... ope of tbc IOVCO!iYC t.:<JfiCCpl ,IS <.>Xprc..,,cci
herein l'<,r cxar:1plc, th<: active ol ihc conl.:m-
platcd cor.1position con simply be mixed 111 !In
aqueous medium to provide ,, mixture ihat can he adminis-
tered to :be affected equine, usually by mollth.
What is claimed is:
10
2. The method of claim 1 wherein the relative weight ratio
of pyrimethamine to sulfonamide is frorr. about 1:15 to
about 1:25.
3. The method of claim 2 wherein the sulfonamide is
sulfadiazine and the relative wetght ratio of pyrimethamine
to sulfadia:Gine is about 1:20.
4. The method of claim 2 wherem the pyrimethamine and
sulfonamide are administered suhstantially wncurrently.
5. The method of claim 2 cvmprising admmtslering the
10
pyrimethamine ar;d the orally.
6. The method of l'iatm 5 com?r!smg the datly admi:Jb--
tration of the pyrimethamine and the sulfonamide
7. The method of dairn 2 cornpns;og aurnini,tc;;rifl5 a
1
, lhcrapcultcltlly ctl'cl:livc amount ol :1 ,-ompnsllion cnmpr"-
ing pyrimethamine and sull'onarntdc, provided that the com-
positioo does not also include trimetho;)rim.
20
8. 'l11e method of c!aim 2 which inhibtls the growth of an
organism irom the genus Sarcocystis.
9. The method of claim 2 which is effective in at least
about 70% of cases.
selected from the group consisting of sulfachlorpyridazine,
sulfadimct hoxine, sulfamerazine, su If a me th a.doe,
sulfamethoxypyridazinc, su!faphcnazole, sulfapyrazine, and

ll. The method of claim 2 in which the is
!'rom lhe group on,i:.:ir.g or nc.:etyl
ll'!llliC t ho)l.ypyrn;.t N -2-lurtnybu: lhtlm id tnr,
.Ill '>ull'Hdoxhe, sulfa:cnc, sulfamcter.
sulfapcrinc, and sulfaphcnuolc
1. A method of treating equine protozoal myclocnccpha
litis (EPM) comprising administering to an equine suspected JS
of sufferi:Jg from EPM therapeutically e!Iective amounts of
pyrimethamine and a sulfonamide, wherein the relative
weight r.1tio of pyrimethamine to the sulfona:nide is from
about 1: lO to about 1 :30; and
12. The method of datm 2 in which the pyrimcthamin<.> is
6dministcrcd in a daily dosage of about 15 to about 30 mg,/kg
of equine.
13. The method of claim 2 further comprising the admin-
istration of folic acid.
14. The method of claim 13 in which the folic acid is
administered daily at a dosage of about 40 mg per 1000-
pound equine.
whertir., when the is sulfadi!tl.lnc, the l'orn- 4n
not conloinlnmct:Jopr!rn in an amount in
lhan a,)out L)i !he W(.:ight o!
Jl'l:H..I!Ct/.1\H,;
15. '!be method of ciaim 2 in which the pynmethaminc
and the sulf6diazine are administered m the form of a solid.
EXHIBITD
' .

Date of Approval: November 5, 2004
FREEDOM OF INFORMATION SUMMARY
NADA 141-240
REBALANCE
Antiprotozoal Oral Suspension
(sulfadiazine and pyrimethamine)
"for the treatment of horses with equine protozoal myeloencephalitis (EPM)
caused by Sarcocystis neurona."
Sponsored by:
Animal Health Pharmaceuticals, LLC
Freedom of Information Summary
NADA 141-240, Page ii
TABLE OF CONTENTS
1. GENERAL INFORMATION .................................................. ..
2. EFFECTIVENESS . .. .. .. . .. .. .. . .. . . .. . . . .. . . . . . . . .. . . .. . .. . . . .. . . . . .. .. . . .. .. . .. 2
a. Dosage Characterization . . . . . . . . . . .. . . . . . . .. . . .. . . . .. .. . .. . . . .. . . . . . . . . . . . . . 2
b. Substantial Evidence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . .. .. . .. . . . .. . . . ..... 2
3. TARGET ANIMAL SAFETY................................................... 9
4. HUMAN SAFETY................................................................ 11
5. AGENCY CONCLUSIONS . . . . . ........ ... . .. . .. .. .. . . .... . .. .. .. . .. . . .. . .. ... 11
6. ATTACHMENTS................................................................ 12
J. GENERAL INFORMATION:
a. File Number:
b. Sponsor:
c. Established Name:
d. Proprietary Name:
e. Dosage Form:
f. How Supplied:
g. How Dispensed:
h. Amount of Active
Ingredients:
NADA 141-240
Animal Health Pharmaceuticals, LLC
1805 Oak Ridge Circle, suite 101
St. Joseph, MO 64506
Drug Labeler Code: 068718
Sulfadiazine and pyrimethamine
REBALANCE Antiprotozoal Oral Suspension
Oral suspension
REBALANCE Antiprotozoal Oral Suspension is supplied
in 946.4 mL (1 qt) bottles.
Rx
Each mL of REBALANCE Antiprotozoal Oral Suspension
contains 250 mg sulfadiazine (as the sodium salt) and
12.5 mg of pyrimethamine.
1. Route of Administration: Oral
j. Species: Equine
k. Recommended Dosage: The recommended dosage is 20 mg/kg sulfadiazine and 1
mg/kg pyrimethamine daily or 4 mL of REBALANCE
Anti protozoal Oral Suspension per 110 lb (50 kg) of body
weight per day. The duration of treatment is dependent upon
clinical response, but the usual treatment regimen ranges from
90 to 270 days. Administer at least one hour prior to feeding
hay or grain.
l. Pharmacological Antiprotozoal
Category:
m. Indications: REBALANCE Antiprotozoal Oral Suspension is
indicated for the treatment of horses with equine
protozoal myeloencephalitis (EPM) caused by
Sarcocystis neurona.
2. EFFECTIVENESS:
a. Dosage Characterization:
NADA 141-240 Page 2
Sulfadiazine (20 mg/kg) and pyrimethamine (1 mglkg) once per day for a minimum
of 90 days is the empirical regimen of the rap( currently recommended in the
scientific literature for the treatment of EPM . Sulfadiazine and pyrimethamine are
two different antimicrobial agents which inhibit folic acid synthesis at two different
sites in the same synthetic pathway. The combination of sulfadiazine and
pyrimethamine is synergistic, with the drug combination having an antiprotozoal
effect. Because of the greater frequency and severity of bone marrow suppression at
the 2X dose level, REBALANCE Antiprotozoal Oral Suspension should be
administered at the labeled lX dose level, sulfadiazine (20 mg/kg) and
pyrimethamine (1 mg/kg) once per day for a minimum of90 days.
b. Substantial Evidence:
(1) Historical Control:
EPM is usually a progressive neurological disease. It has been estimated that up
to 55 to 65%
2
of horses respond favorably to treatment. However, it is further
estimated that a small percentage (no more than 1 0%) of treated horses recover
completely. One of the most important points to consider is that EPM produces a
highly variable clinical disease. Historical controls were used in the field studies
because, without treatment, EPM is usually a progressive disease. At the time
these studies were conducted, there was no FDA approved treatment for EPM.
The use of historical controls in the evaluation of compounds for effectiveness
is described in 21 CFR 514.117(b)(4)(iv).
(2) Field Study: Clinical Field Effectiveness and Safety ofDaily Pyrimethamine
and Sulfadiazine Oral Suspension in Horses Affected with EPM
(a) Type of Study:
This study was conducted as a multi-site, randomized field effectiveness
evaluation oftwo dose levels of REBALANCE Antiprotozoal Oral
Suspension. The two dose levels (IX) 20 mg/kg sulfadiazine and 1 mg/kg
pyrimethamine and (2X) 40 mg/kg sulfadiazine and 2 mg/kg pyrimetramine
were administered daily for a minimum of 90 days.
(b) Investigators:
1
MacKay RJ, Granstrom DE, Saville WJ, Reed SM. Equine Protozoal Myeloencephalitis: Veterinary Clinics
of North America/Equine Practice, 2000:16:405-425.
2
Granstrom DE. Understanding Equine Protozoal Myeloencephalitis: Your Guide to Horse Health Care and
Management. Lexington: The Blood-Horse Inc., 1997:10.
Dr. Laurie A. Beard
Ohio State University
Veterinary Teaching Hospital
Columbus, OH
Dr. Deme M. Erickson
Rochester Equine Clinic
Rochester, NH
Dr. Philip J. Johnson
University of Missouri
College ofVeterinary Medicine
Columbia, MO
Dr. John M. Leonard
Fox Run Equine Center
Apollo, PA
(c) Study Design:
NADA 141-240 Page 3
Dr. Stephen M. Reed
Ohio State University
Columbus, OH
Dr. Bonnie Rush
Kansas State University
Manhattan, KS
Dr. Harold C. Schott
Michigan State University
College of Veterinary Medicine
East Lansing, MI
Dr. Corinne R. Sweeney
University of Pennsylvania
School ofVeterinary Medicine
New Bolton Center
Kennett Square, P A
1 Purpose: This study was designed to evaluate the clinical
effectiveness and safety of sulfadiazine and pyrimethamine for the
treatment of horses with EPM at a dosage of 20 mg/kg sulfadiazine
and 1 mg/kg pyrimethamine or 40 mg/kg sulfadiazine and 2 mg/kg
pyrimethamine administered in a daily oral dosage for a minimum of
90 days .
.f. Test Animals: There were 97 horses enrolled in this study, consisting
of 34 females, 13 males and 50 geldings ranging from nine months to
32 years of age. Seventy-two percent of the horses were
Thoroughbreds, Standardbreds and Quarter Horses, with the
remainder represented by Tennessee Walker Horses, Appaloosas,
Arabians and mixed breeds.
Enrollment Criteria: Initial selection of the animals into the field
effectiveness study was based upon a qualifying physical
examination and on a clinical neurological examination. Blood and
cerebrospinal fluid (CSF) samples were collected for determination of
serum and CSF serological status for EPM (Western Blot Test),

t
Freedom oflnformation Summary
NADA 141-240 Page 4
albumin quotient (AQ) and immunoglobulin 0 (IgG) index. In
addition, blood samples were collected for complete blood count
(CBC) and serum chemistry profiJe to assess the overall health of the
animals prior to initiating treatment.
Subsequently, a diagnosis ofEPM was confirmed by a positive
Western Blot Test for Sarcocystis neurona on CSF and clinical
signs compatible with EPM. Animals were admitted to the study
prior to receipt of results ofCSF Western Blot Test, but eligibility
was revoked upon receipt of a negative CSF Western Blot Test. The
severity of the neurological deficit was determined by the overall
neurological dysfunction (OND) score.
Neurologic Grading Score for OND:
0 ==clinically normal. No detectable dysfunction.
I = slight deficit. Dysfunction barely perceptible.
2 = moderate deficit. Dysfunction easily detectable.
3 = marked deficit. Dysfunction strikingly conspicuous.
4 = severe deficit. Profound dysfunction.
5 = recumbent.
i Exclusion Criteria: Animals outside of study specifications, such as
pregnant mares, horses with clinical histories incompatible with EPM
diagnosis, or CSF Western Blot Test negative serological status, were
excluded from the field effectiveness study. Animals whose
owners/authorized agents had not signed the informed consent
document were also excluded from the study. Animals which had
condition(s) other than EPM that might interfere with the clinical
determination of severity of the neurological deficit caused by EPM
and the response to treatment, were also excluded from the study.
Animals which had been treated for EPM condition for more than 30
days immediately prior to admittance to this clinical effectiveness
study were also excluded from the study.
Treatment Groups and Controls: Ninety-seven (97) horses met the
clinical effectiveness study entrance criteria. Each horse admitted to
the study had a baseline clinical evaluation consisting of a clinical
description and characterization of myoneural abnormalities, a
videotape recording of the neurological deficit, a physical
examination, a CBC, a serum chemistry analysis, a CSF and a serum
Western Blot Test and a determination ofCSF indices (AQ and IgG
index protein electrophoresis) prior to assignment to a treatment
group.
Treatment dosage of sulfadiazine and pyrimethamine (IX and 2X)
,
,

NADA 141-240 Page 5
was predetermined by a randomized treatment schedule. Forty-eight
(48) horses were assigned to the lX treatment group (20 mg/kg
sulfadiazine and l mg/kg pyrimethamine); forty-nine horses were
assigned to the 2X treatment group (40 mg/kg sulfadiazine and 2
mg/kg pyrimethamine). The scheduled dosage regimen was for daily
oral administration for a duration of90 to 180 days. In 14 horses, the
duration of administration exceeded 180 days, with the duration of
administration ranging from 195 to 270 days.
Dosage Form: The oral suspension formulation used during this study
was identical to the product intended for marketing. REBALANCE
Antiprotozoal Oral Suspension contains 250 mg/mL sulfadiazine (as
the sodium salt) and 12.5 mg/mL pyrimethamine.
1 Route of Administration: REBALANCE Antiprotozoal Oral
Suspension was administered by the oral route via syringe.
Dose, Frequency and Duration: The horses were dosed with either IX
(20 mg/kg sulfadiazine and I mg/kg pyrimethamine) or 2X (40 mg/kg
sulfadiazine and 2 mg/kg pyrimethamine) daily for a duration of90 to
270 days. Horses were dosed at least one hour prior to feeding hay or
grain .
.2 Treatment Success: The primary effectiveness variables for the
determination of response to test article treatment were the CSF
Western Blot Test results and the overall assessment of neurological
dysfunction (OND score). A horse was considered a success if any
of the following criteria applied at the time the horse was evaluated:
Negative CSF Western Blot Test and clinical neurological
improvement (one or more grade improvement in OND score)
Negative CSF Western Blot Test and no clinical neurological
improvement (zero or less improvement in OND score)
Positive CSF Western Blot Test and marked clinical neurological
improvement (two or more grades improvement in OND score)
All neurological examinations were videotaped. In order to
corroborate the investigators' OND scores, independent experts were
asked to view the videotapes and confirm that the horses that were
deemed to be clinical successes by the investigators appeared to
improve on videotape. If the independent experts agreed that the horse
showed improvement, the horse was considered a corroborated
success.
(d) Results:
.

NADA 141-240 Page 6
Forty-eight horses were assigned to the 20 mg sulfadiazine/kg and 1 mg
pyrimethamine/kg dose of REBALANCE Antiprotozoal Oral Suspension
(1 X treatment group). The final database consisted of 26 horses treated
at the 1X dose, the other 22 horses in the IX group failing to complete the
study. Day 0 was the day the horse was first administered test
article. Evaluations were made every 30 days. All horses were treated
with drug at least 90 days.
Table 1. Summary of Effectiveness Outcomes for 26 horses treated at the lX dose.
Day of 30 60 90 120 150 180 >210 Totals
Evaluation
CSF Successes
- -
2 1 2
-
.
5
OND Successes 1 1 2 3 2 1 l 11
Uncorroborated
- - - -
-
1 l 2
OND Successes
Failures
- - -
2 1 3 4 10
OND Successes 1 1 4 4 4 1 1 16
&CSF
Successes
Corroborated 1 1 4 4 4 0 0 14
OND Successes
&CSF
Successes
Based on the clinical investigator's evaluations and the results ofthe CSF Western
Blot Analysis, 16/26 (61.5%) of horses treated at the IX dose were successes. Based
on the corroborated clinical investigator's evaluation and the results of the CSF
Western Blot Analysis, 14/26 (53.8%) of horses treated at the lX dose were
successes. The 95% Blyth-Still-Casella confidence interval for the cumulative
percent of corroborated successes is (33.4%, 71.8%).
(e) Adverse Reactions:
Adverse reactions pertaining to bone marrow suppression were two or more times
more frequent in the 2X treatment group than the IX treatment group. Adverse
reactions were categorized under the following categories: bone marrow, appetite,
gastrointestinal, integument, treatment crisis and unusual daily observations.
Although 97 horses were enrolled in the study (48 in the 1X treatment group, 49
in the 2X treatment group), only 75 horses were administered the drug in the
two treatment groups for a duration of at least 90 days, thus a total of 75 horses
were evaluated for adverse reactions. Adverse reactions were evaluated in 37
horses treated with the IX dosage, which included 5,910 daily observations, and
38 horses treated with the 2X dosage, which included 6,210 daily observations.
t
NADA 141-240 Page 7
1 Bone Marrow: Bone marrow suppression due to test article administration
caused overall anemia (classification of anemia based on RBC, Hgb,
PCV/HCT variables) in 12% of the scheduled observations in the lX group
and 21% of the scheduled observations in the 2X group. In the 37 clinical
cases that were treated with lX dose of test article 90 or more days, anemia
was noted in 22%, leukopenia in 19%, neutropenia in 5% and thrombocyto-
penia in 3% of the cases. Similarly, in the 38 clinical cases in the 2X
treatment group, anemia was noted in 58%, leukopenia in 55%, neutropenia
in 29% and thrombocytopenia in 5% ofthe cases. The incidence of bone
marrow suppression in the 2X treatment group was two or more times that
ofthe lX treatment group and the degree of suppression was more serious
(mild to severe vs. mild to moderate). Because of these blood dyscrasias,
test article was interrupted over four times more often in horses treated at
the 2X dosage than those treated at IX, although both groups were off
treatment for about the same amount of time ( ~ 2 0 of the treatment
period). In some instances of bone marrow suppression, diet was
supplemented with folinic acid to aid in recovery of the bone marrow.
Interruption of test article administration with or without folinic acid
supplementation proved adequate in preventing any detrimental effects to
the overall health and well being of the test animals. Blood counts were not
low enough for a long enough period of time to allow development of
clinical signs.
Z Appetite: Anorexia was reported in 0.24% of the daily observations for this
category in the lX. treatment group (two out of37 horses) and in 0.03% of
the daily observations for the 2X treatment group (one out of38 horses).
The one horse in the 2X treatment group exhibiting anorexia had a
concurrent fever associated with liver disease/cholestasis and/or
enterotoxemia (hepatocellular disease). Decreased appetite (off feed) was
reported in 0.10% of the daily observations for the 1 X treatment group (one
out of 37 horses) and in 0.05% of the daily observations for the 2X
treatment group (one out of 38 horses).
l Gastrointestinal: Loose stools were observed in three out of37 horses in the
1 X treatment group and five out of 38 horses in the 2X treatment group.
Twenty-four of the 26 loose stool observations in both treatment groups
occurred between Day 0 and Day 30. Fifteen loose stool observations
occurred in one horse in the lX treatment group between Day 0 and Day
30. Diarrhea was observed in one out of 38 cases in the 2X treatment group
(one observation on Day 4).
1: Integument: Urticaria was observed in one out of37 horses in the lX
treatment group and in two out of 38 horses in the 2X treatment group.
One of the horses received conservative topical treatment and two horses
received no treatment. The urticaria resolved without sequelae.
NADA 141-240 Page 8
2 Treatment Crisis (marked worsening of the neurological condition during
treatment believed to be due to an inflammatory reaction in the CNS to the
dead/dying protozoan organisms): One horse in the IX treatment group
became progressively more neurologic and recumbent after 99 days of
treatment. The worsening of the neurologic signs was assumed to be due to
inflammation in the central nervous system associated with dead or dying
protozoa. The horse was euthanized on test day 114 .
.2 Unusual Daily Observations: The following daily observations were also
noted. Lethargy/mild depression was observed in five horses; seizure
occurred in one horse; mild colic was observed once in three individual
horses; elevated liver enzymes associated with acute onset of hepatocellular
disease was observed in one horse; increased urination/defecation was
observed in one horse; fever was observed in four cases (two with upper
respiratory infection, one with hepatocellular disease and one of unknown
etiology); neutrophilia associated with inflammation, infection, and/or
stress was observed in five horses; leukocytosis associated with upper
respiratory infection or hepatocellular disease was observed in two horses;
and itchiness/delayed shedding was observed in one horse.
1 Serum Chemistry: There was no test article affect on any of the clinical
chemistry variables.
(f) Conclusions:
Based on the clinical investigator's evaluations and the results ofthe CSF
Western Blot Analysis, 16/26 (61.5%) of horses treated at the IX dose were
successes. Based on the corroborated clinical investigator's evaluation and the
results of the CSF Western Blot Analysis, 14/26 (53.8%) of horses treated at
the 1 X dose were successes. The total number of horses that became CSF
Western Blot negative was five out of 26 or 19.2%.
REBALANCE Antiprotozoal Oral Suspension is effective for the treatment of
horses with equine protozoal myeloencephalitis (EPM) caused by Sarcocystis
neurona.
,..
I
3. TARGET ANIMAL SAFETY:
(a) Type of Study: Toxicity
(b) Investigator and Trial Location:
Dr. Steven G. Kamerling
NADA 141-240 Page 9
Department Of Physiology, Pharmacology and Toxicology
School of Veterinary Medicine
Louisiana State University
Baton Rouge, LA
(c) Study Design
1 Compliance: This study was conducted in compliance with the FDA Good
Laboratory Practice Standards, 21 CFR 58 .
.f. Purpose: The purpose of the study was to evaluate the safety of REBALANCE
Antiprotozoal Oral Suspension in horses .
.l Test Animals: Fourteen healthy Thoroughbred and Quarter Horse test animals
(seven males and seven females), ranging in age from three to 16 years .
.1. Dosage Form: The test article was an oral suspension .
.2_ Route of Administration: The test article was administered orally.
. Dosage, Frequency and Duration of Treatment: Ten horses (five males and five
females) were administered REBALANCE Antiprotozoal Oral Suspension at
a dosage of 8 mL/50 kg (11 0 lbs) a day (2X the labeled dose) for 92 days.
Treatments were given at least one hour prior to feeding of hay and grain.
1 Controls: Four horses (two males and two females) were not treated.
Evaluation of Variables: The health of each animal was evaluated daily. A
complete physical examination was conducted and blood was. drawn for CBC
and serum chemistry analyses three times prior to treatment on test days minus
14 and minus 7 for study eligibility and on Day 0 (immediately prior to start of
treatment). The average of the two baseline measurements on test days minus
14 and minus 7 were used as the covariate for each response variable. The
physical examinations and blood analyses were repeated at 14-day intervals
during treatment and 14 and 29 days following the end of treatment (total of
eight physical examination/blood sample observations/analyses per animal
during/following treatment).
"'\
f l
NADA 141-240 Page 10
,2 Statistical Methods: CBC and clinical chemistry variables were analyzed using
analysis of covariance. Treatment and treatment by time interactions were
considered significant if their p-values were less than or equal to 0.10.
(d) Results:
1 Physical Examinations: The observations from biweekly physical examinations
were not associated with any clinically significant condition in either group.
Complete Blood Count (CBC): CBC variables were analyzed using repeated
measures analysis of covariance. Both the treated and control groups
experienced a decline in RBC starting on Day 70; however, the treated group
was statistically significantly lower (p,S 0.1 0) than the control group on Days
105 and 120. Hgb also decreased in both groups; however, it was statistically
significantly lower (p.S 0.10) in the treated group on Days 28 and 120. PCV
and HCT decreased along with the RBC; however, there were no statistically
significant differences between treated and control groups. MCV values
remained within normal limits for both groups; however, MCV values were
slightly elevated in the treated group on Days 70, 84, 105 and 120. Despite the
findings of the CBC, there were no clinical signs of anemia observed in either
group. Twenty-nine days after cessation of treatment, all values returned to or
above baseline levels in both groups. There were no clinically significant
changes in white blood cells.
l Clinical Chemistry: Most serum chemistry variables remained within normal
limits throughout the study in both treated and untreated groups. There was
considerable variation between groups and from baseline values in both groups,
e.g., creatinine, alkaline phosphatase (ALP), gamma glutamyl transferase
(GGT), total protein, globulin and albumin. Elevated ALP values were
observed in three of the ten treated horses. These values were slightly above
the upper end of the normal range on study days 84 and 105 .
. Daily Observations: Daily animal care observations indicated that the test
article was well tolerated for the duration of the treatment period. The most
significant observation was the transient appearance of loose stools in both the
treated and untreated groups, although the frequency of occurrence was greater
in the treated group. Diarrhea was infrequently observed in the treated group
and not observed in the untreated group. At no time during the study was the
occurrence of loose stools or diarrhea worthy of medical intervention and all
cases resolved without sequelae.
2. Appetite: Depressed appetite occurred infrequently in all but one of the treated
horses during the study period (affected horses had a depressed appetite for one
or two days during the 92-day treatment period). Depressed appetite was not
observed in the untreated group. In one of the treated horses, depressed
appetite progressed to anorexia. Daily ration for the anorectic horse was
changed from a pelleted ration to a sweet feed ration and appetite was restored .
. Unusual Observations: One transient case of urticaria in a treated horse was
observed on Day 41. This resolved without treatment within 24 hours.
Another treated horse became acutely ataxic on Day 88 and died within two
hours. Post-mortem examination revealed focal hemorrhage ofthe brainstem
and cerebellum, a lesion consistent with the clinical signs observed prior to
death. The precise cause of this cerebrovascular accident was not determined.
This horse was also reported to have oral ulcers on Day 45 of the study.
(e) Conclusions:
REBALANCE Antiprotozoal Oral Suspension, administered at 2X the
recommended label dose for 92 days resulted in clinical signs of toxicity
including: partial to complete anorexia, loose stools and diarrhea, mild to
moderate anemia and elevated ALP levels. None of these adverse effects
required medical intervention.
4. HUMAN SAFETY:
This drug is intended for use in horses, which are non-food animals. Because this new
animal drug is not intended for use in food-producing animals, data on human safety
pertaining to drug residues in food were not required for approval of this NADA.
Human Warnings are provided on the product label as follows: "For use in horses only.
Do not use in horses intended for human consumption. Not for human use. Keep out of
the reach of children."
5. AGENCY CONCLUSIONS:
The data submitted in support of this NADA satisfy the requirements of section 512 of
the Federal Food, Drug, and Cosmetic Act and 21 CFR Part 514 of the implementing
regulations. The data demonstrate that REBALANCE Antiprotozoal Oral Suspension,
when administered under labeled conditions is safe and effective for the treatment of
horses with equine protozoal myeloencephalitis (EPM) caused by Sarcocystis neurona.
The drug is restricted to use by or on the order of a licensed veterinarian because
professional expertise is critical for the diagnosis of equine protozoal myeloencephalitis
in horses. The safe use of this product should also be monitored by the veterinarian.
Under section 512(c)(2)(F)(ii) ofthe Federal Food, Drug, and Cosmetic Act, this
approval qualifies for THREE years of marketing exclusivity beginning on the date of
the approval.
I
REBALANCE Antiprotozoal Oral Suspension is protected under the following
U.S. patent numbers:
!J.S. Patent Number
5,747.476
6,255,308
6,448,252
6. ATTACHMENTS:
Date of Expiration
July 17, 2016
July 17,2016
July 17,2016
Facsimile labeling is attached as indicated below:
a. Bottle Label
b. Package Onsert

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