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What is ALL?
Acute lymphoblastic leukemia (ALL) is a form of leukemia in which immature white blood cells (blasts) become malignant and proliferate excessively in the bone marrow.
High number of blasts crowd healthy mature cells and prevents their proliferation.
Blasts can enter into the periphery and become metastatic and may form solid tumors in other tissues. CNS Organs
Testicles
Chest
Leukopenia
Fatigue
Frequent infections
Fevers Chills
Weakness
Bleeding Bruising Swollen/Bleeding gums Petechia Loss of Apetite Bone/Joint pain Weight loss
Thrombocytopenia
Other
Anemia
Diagnosis of ALL
The most common diagnostic tool is a blood smear and total count
If ALL is suspected, a bone marrow biopsy and lumbar puncture may be performed to see the extent of the cancer.
The American Cancer Society predicts that there will be 5,730 new cases of ALL reported in 2011. 75% of leukemia cases in children and adolescents (0-19) are ALL the most common cancer in children under 20. Treatment of the disease has improved but Lasparaginase (L-ASNase) continues to have a high rate of immunogenicity.
L-ASNases
Interest in this enzyme as a therapeutic began in the 1960s when guinea pig serum showed to have anticancer properties. It was later discovered that guinea pig serum has heightened levels of circulating L-ASNase. L-ASNases have been found in many bacteria, yeast, molds, plants and in the plasma of certain vertebrates.
Structure of L-ASNases
Native form produced from E. coli (Elspar) Native form produced from E. chrysanthemi Polyethylene glycol (PEG) modified E. coli L-ASNase (Oncaspar)
PEGylation
Refers to the covalent addition of at least one PEG chain to a molecule. PEGylation improves protein drugs but may be associated with loss of bioactivity.
Mechanism Continued
L-ASNases catalyze the hydrolysis of L-asparagine (Asn) to aspartic acid and ammonia. Healthy human cells have sufficient concentrations of asparagine synthase. They do not rely solely on extracellular Asn for proper function. Leukemic blasts do not have sufficient concentrations of the enzyme and cannot up regulate its expression when extracellular [Asn] gets low. Lack of Asn inhibits protein synthesis in blasts and causes them to undergo apoptosis (cell cycle arrest at G1 phase).
Oncaspar/pegaspargase
Oncaspar is a PEGylated version of the native E. coli LASNase. L-ASNase is a foreign protein and commonly induces the production of antibodies (58%) and sometimes severe allergic reactions (24%, 29%) Oncaspar has 5000-dalton units of monomethoxypolyethylene glycol conjugated to the enzyme. The PEG groups on Oncaspar
Reduce its immunogenicity Lower incidence of silent antibodies Increase t1/2 of the drug reduce antibody mediate rapid clearance Maintains the safety profile of native L-ASNase More convenient lower doses and less frequent administration
Oncaspar/pegaspargase
Oncaspar is manufactured by Sigma-Tau who acquired it from Enzon. The native E. coli L-ASNase is produced and isolated from E. coli and provided to Sigma-Tau by Lundbeck (makers of Elpar). The enzyme is then PEGylated using the Enzons Customized Linker Technology platform Enzon is payed a 5-10% royalty for the sale of Oncaspar.
Oncaspar first approved by the FDA in February of 1994 for the treatment of patients with ALL who suffered sever immune responses to native E. coli L-ASNase. The drug later received FDA approval in July of 2006 for first-line treatment of ALL in multiagent chemotherapy regimens.
Patients
31 individuals treated
Treatment
Patients
Drug was delivered IV over 1 hour for every 14 days Results based on 27 evaluable patients
Disappearance of PEG-L-ASNase from plasma with an average t1/2 of 357 hours* (14.8 days) biweekly administration.
Concentration of PEG-LASNase in plasma, after the first IV infusion and at day 14, proved to be proportional to the administered dose.
Rate of total clearance of PEG-L-ASNase found to be 128 mL/m2/day compared to E. coli L-ASNase 2196 mL/m2/day.
1 at 500 IU/m2
1 at 2000 IU/m2
1 at 4000 IU/m2
Patients
148 patients (93 M, 55 F) 20 years old B-precursor ALL
Treatment
Reinduction with:
PEG-ASNase either weekly or biweekly (at random) at 2500 IU/m2 (IM) Combination drugs following standard treatment administered (doxorubicin, prednisone, and vincristine)
Evaluations
2 times per week. Serum alanine aminotransferase (ALT), total and direct bilirubin, albumin, glucose, amylase, lipase, and plasma fibrinogen days 1, 15, and 29. Bone marrow aspiration days 15 and 29 Serum L-ASNase concentration weekly on days 8, 15, 22, and 29 Serum samples for E coli L-ASNase and PEG-LASNase antibodies weekly on days 8, 15, 22, and 29
CBC
Analysis
L-ASNase enzyme activity measured by coupling L-ASNase activity to oxidation of NADH to NAD+ (by alphaketoglutarate/oxaloacetate) and reading reactions at 340nm.
Serum anti-L-ASNase antibodies assayed using an antibody capture ELISA.
Weekly drug administration tended to show higher incidence of complete remission and lower incidence or disease resistance.
Infections were common and resulted in the death of 4 patients. Unrelated to drug
High titer of antibodies against E. coli ASNase or PEG-ASNase correlated with low ASNase levels.
Adverse events were as common as the 2% to 3% incidence documented in other studies using E. coli ASNase in treatment. More frequent administration of the drug maintained higher PEG-ASNase concentrations in plasma which is associated with lower antibodies.
Patients
Maintenance phase
Duration was 2 (girls) and 3 (boys) years
Patients also received the standard combination chemotherapy drugs (Vincristine, Prednisone, Cytarabine, Methotrexate, etc) Patients were randomly assigned to receive either
Treatment
4 weeks of induction 4 weeks of consolidation 2 8-week interim maintenance phases 2 8-week delayed intensification (DI) phases
2500 IU/m2 of Oncaspar intramuscularly (IM) on day 3 of induction and each DI phase or 6000 IU/m2 of native E. coli ASNase IM 3 times per week, for 9 doses in induction, and 6 doses in each DI phase.
Patient Monitoring
Physical exam
Blood and urine tests
Serum anti-L-ASNase antibodies assayed using a modified indirect solid-phase ELISA. AAs asparagine, aspartic acid glutamic acid and glutamine quantified via high-performance liquid chromatography.
Analysis
L-ASNase enzyme activity measured by quantifying ammonia produced from ASN read by ELISA.
Oncaspar showed better clearance of blasts in ALL infected bone marrow than the E. coli L-ASNase.
Oncaspar Day 7 Oncaspar Day 14 E. coli L-ASNase E. coli L-ASNase Day 7 Day 14
M1 (<5% blasts)
M2 (5%-25% blasts)
M3 (>25% blasts)
Oncaspar showed better clearance of blasts in ALL infected bone marrow than the E. coli L-ASNase.
The study sought to show a decrease in anti-L-ASNase antibodies when Oncaspar was used. This graph shows that pegaspargase significantly reduced the amount of antibodies produced.
*Ratio reflects the amount of antibodies in patients sera over the amount of antibodies in healthy individuals sera (control)
Oncaspar has a longer t1/2 than E. coli L-ASNase (5.5 vs. 1.1 days).
Greater L-ASNase concentration though dosage and frequency of administration was lower.
Oncaspar showed similar correlation between clearance of AAs in serum and CSF when compared to E. coli L-ASNase.
Note:
Gln
Gln
Asn
Asn
Production of anti-L-ASNase antibodies cause severe/life threatening allergic reactions and can silently decrease the efficacy of the drug, preventing its further use. Oncaspar, when compared to E. coli LASNase, showed (why its better):
Lower production of allergic response initiating and enzyme inactivating antibodies Better clearance of blasts from infected patients bone marrow More persistent and higher L-ASNase activity Similar, if not better, toxicity and efficacy Longer t1/2 (by 4+ days) The need for less frequent administration and lower doses of the drug 1 administration can replace 6 to 9 administrations of E. coli L-ASNase. Better EFS probability (88% vs. 85%)
Pharmacoeconomic component of the trial showed patient costs of Oncaspar was similar* to E. coli L-ASNase.
*Later studies claim that the overall costs associated with treatment using pegaspargase would be considerably less compared to therapies using conventional L-
Industrial Perspective
Sigma Tau has another ALL drug in Phase III clinical trials called EZN-2285 (calasparagase pegol) Future Treatments:
Mesenchymal
treatment Modifying ASNase to alter glutimase activity reduce side effects but have greater effect against cancer cells Modifications:
Entrapment
of enzyme in liposomes/microcapsules/RBC Covalent coupling to macromolecules (dextran, albumin, ) Immobilization on polyacrylamide or agarose
$50 million dollars to avoid drug shortage Cost of Oncaspar jumped from $2,625 to $5,670 per vial Could be as high as $40k for full treatment. With government programs like Medicaid $70/$490
is estimated at $190 per vial Administration is more frequent Oncaspar shown to be equivalent to 9 doses Considering costs of complications and office visits it has been estimated to not save much, if any, money.
patients by reimbursing patients copays 100% No limitations on family income or financial status S.O.S program provides Oncaspar free of charge to eligible patients who may not have insurance coverage or the ability to pay.
Patents
Patent Number Date
Patent Title
Inventors/Assignee
Pierce Chemical Company
Davis; Frank F. The United States of America as represented by the Department of Health Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo Enzon, Inc.
Sources
American Cancer Society. "Cancer Facts & Figures 2011." American Cancer Society - Information and Resources for Cancer. Web. 07 Oct. 2011. <http://www.cancer.org/Research/CancerFactsFigures/CancerFactsFigures/cancer-facts-figures-2011>.
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