65

Structure Activity Relations of Polyfunctional Diterpenes of the Tigliane Type, VI

(8) Mann,J. D., E. A. Edge,J. E. Lancaster,Blyth,K.(1980)N. Z.J.

agric. Res. 23,361.
(9) Sinicin, G. S. (1963)Tr. Inst. Bot. Akad. Nauk. Kaz. SSR. 17, 153.
Hort. Abstr. 35,6374(1965).

(10) Balcar-Skrzydlewska, E., Zelecha, M. (1964) Biul. Inst. Rosl.

Lecz. 10, 159. Hort. Abstr. 36,5266(1966).
(11) Slavik, B. (Ed.): Methods of Studying Plant Water Relations, 450
pp., Berlin, 1974, Springer Verlag.
(12) Arnon, D. I. (1949) P1. Physiol. 24,1.
(13) Gairon, S., Hadas, A. in: Arid Zone Irrigation Ecological Studies
5,215, New York, 1973, Springer-Verlag.
(14) Birner, J. J. (1969) Pharm. Sci. 58,258.
(15) Weissenberg, M.: Unpublished results.
(16) Hellebust,J. A. (1976) A. Rev. P1. Physiol. 27,485.

(17) Turner, N. C. and M. M. Jones, in Turner, N. C. and Kramer, P. J.

(Eds.): Adaptation of Plant to Water and High Temperature

Stress, p. 482, New York, N. Y., 1980, J. Wiley and Sons.
(18) Turner, N. C.,in Musell, H. and R. C. Staples (Eds.): Stress Physiology in Crop Plants, pp. 343372, New York, N. Y., 1979, J.
Wiley and Sons.
(19) Turner, N. C. and P. J. Kramer (Eds.): Adaptation of Plants to
Water and High Temperature Stress, pp. 720, New York, N. Y.,
1980, J. Wiley and Sons.

(20) Fischer, R. A., Maurer, R. (1978) Aust. J. agric. Res. 29, 897.
(21) Clarckson, N.M., Rusell, J. S. (1976) Aust. J. agric. Res. 27,227.
(22) Yaniv, Z., M. Weissenberg, Palevitch, D. (1983) Acta Hort. 132,
217.

(23) Yaniv, Z., M. Weissenberg, D. Palevitch, Levy, A. (1981) Planta

Med. 42,303.
(24) Miller, L. M. F., Davies, M. E. (1979)J. Linn. Soc. Symp. Ser. 7,
231.

Structure Activity Relations of Polyfunctional Diterpenes

of the Tigliane Type, V11
Irritant and tumor promoting activities of semisynthetic mono and diesters of 12-deoxyphorbol
S. Zayed2'4, B. Sorg4 and E. Hecker3'4

Abstract: A method for the isolation and preparation of 12-deoxyphorbol (1) from Euphorbium resin (Euphorbia resinifera Berg) was established to provide substantial amounts of 1-esters for bioassays. Acylation of 1 yielded 12-deoxyphorbol-13,20-diesters (2, esters with acids
CH3(CH2)COOH, n = 0,4,8, 12, 16,20; and with benzoic, oleic, elaidic and linoleic acid). Upon mild transesterification the 13,20-diesters 2
yielded the 13-monoesters 3. Both ester types were tested for their irritant activities, the 13-monoesters were also assayed for their tumor-

promoting activity. A dependance of both of these activities on the

chain length of the acyl residues is noticeable. For both probably a broad maximum exists around the tetradecanoate.

Introduction

experiment the 12-deoxyphorbol-13-tetradecanoate (3) appeared to exhibit a promoting activity (3) about as strong as that of
the well known standard promoter 12-O-tetradecanoylphorbol-13-acetate [Croton oil factor A1, TPA (5)]. To study structure activity relationships of 12-deoxyphorbol esters (6) in a systematic manner, a method to provide substantial amounts of
the diterpene parent 12-deoxyphorbol (1) from the easily ac-

cessible Euphorbium resin was developed and a variety of

13,20-diesters (2) and 13-monoesters (3) was prepared and
their biological activities tested.

18.

-0R2

12

13

Within the last ten years numerous 13-mono-(3) and 13,20-di-

esters (2) of the diterpene alcohol 12-deoxyphorbol (1, for

structural formulas see Fig. 1) were isolated from various Eu-

phorbiaceae and Thymeleaeceae species [for reviews see:

(1,2)]. As far as tested, these esters showed more or less pronounced irritant activity on the mouse ear (24). Moreover,
some of the 13-monoesters tested also showed weak tumorpromoting activity on the back skin of mice (3,4). In a singular

Vth communication: Marston, A., Hecker, E. (1983) Z. Naturforsch. 38b, 1015

2

O 6/

1: R1=R2=I-I
2: R'=R2=acyl
3: R1=H,R2=acyl

CH2OR1
20

Fig. 1 Structural formulas

of the 12-deoxyphorbol esters prepared

Materials and Methods

Permanent adress: National Research Centre, Dokki, Kairo, Egypt

To whom reprint requests should be sent

Institut fr Biochemie, Deutsches Krebsforschungszentrum, D-6900
Heidelberg 1, Im Neuenheimer FeId 280

Melting points are uncorrected. The IR-spectra were carried out on a

Perkin-Elmer spectral photometer 521. UV-spectra were registered on
a Beckman DK-2a far UV-spectrometer. mass spectra (MS) were mea-

Downloaded by: NYU. Copyrighted material.

(5) Foldesi, D., Lang, T., Kovacs, T. (1969) Herba Hung. 8,49.
(6) Fryer, R. F. (1972) Proc. Agron. Soc. N. Z. 2,73.
(7) Bernath, J., Foldesi, D. (1972) Herba Hung. 11,55.

Planta Medica 1984

66
sured with a Dupont CEC-21-110 B and 60 MHZ 'H-NMR-spectra
were measured with a Jeol HL C-60 spectrometer. The NMR-spectra
were recorded with tetramethylsilane (6 = 0.00 ppm) as internal stan-

C). Yields, R1 values and mass spectral data of all 13,20-diesters 2 prepared are compiled in Table I. For UV, IR and 'H-NMR data compare
the 13 ,20-diacetate (7) as a representative.

dard. The Euphorbium (Euphorbia resinifera Berg) resin was obtained

matography, Merck silica gel 0.050.2mm deactivated with 13% water or Polyamide (Woelm) were used. Spots were detected under UVlight (254 nm) and made visible by spraying with vanillin/sulfuric acid
(7).
Working-up procedure
The reaction mixture was poured into twice its volume of water and
stirred for 1 h. After extraction with ethyl acetate the organic layer was
washed with 2 M HC1, then with 5% aqueous KHCO3 and finally with
water. Following drying over sodium sulphate the solvent was removed
under reduced pressure.

Isolation and preparation of 12-deoxyphorbol (1) from Euphorbium

resin
Euphorbium resin (1kg) was shaken with 51 of methanol. Following removal of the insoluble residue by filtration, the filtrate was added to a
mixture of 250 ml of water and 2 I of petroleum ether (b.p. 6080C).

After 1 h of shaking the petroleum ether layer was discarded. After

evaporation, the water methanol phase afforded 300 g of a resinous residue, which was suspended in 600 ml of ethyl acetate and filtered over
a silica gel (1 kg) column using ethyl acetate as mobile phase. After evaporation, the filtrate gave 130 g of a resinous material, which was chromatographed on a silica gel (1.5 kg) column using a gradient elution
with ethyl acetate/benzene mixtures. The fraction containing esters of

13-Monoesters 3 by partial transesterification of 13,20-diesters 2.

(a) 12-Deoxyphorbol-13-acetate. To a solution of 12-deoxyphorbol13,20-diacetate (2) (470 mg) in methanol (300 ml), 70% HCIO4 (0.4
ml) was added. The mixture was allowed to stand under nitrogen for 24
h. Sodium acetate (600 mg) was added, methanol was removed in vacuum and the residue was extracted with ethyl acetate. After drying and
evaporation of the solvent the crude product was purified by TLC (1/15
CH3OHJCH2CI2); yield: 210mg 13-acetate 3. MS: m/e 390 (Mt). 'HNMR (d5-pyridine): 1-H: 7.74 (m); 7.H: 6.1 (d, J78 = 5 Hz); 20-H2:
4.32 (s); 10-H: 3.7 (m); 8-H: 3.6 (m); 5-H2: 3.05 (s); 19-H3: 1.7 (m);
acetyl: 2.04; OH (exchangeable with D2O): 7.82 and 4.88 ppm (2 s). IR
(KBr): 3440 (OH), 1700 (CO), 1620 cm' (C = C). UV (methanol):
?.,,, (a) = 234 (5240), 333 nm (70).
(b) Monoesters 3 other than 13-acetate. To a solution of the 13,20diester 2(1.5 mmole in 10 ml of methanol), sodium methoxide in methanol (0.75 mmole, used as 10_2 M solution) was added. The reaction
mixture was kept at 5 C and tested by TLC (1/10 CH3OWCH2CI2)
every 30 mm in order to determine the time of optimal partial transesterification. The reaction was then quenched with acetic acid and the solvent was removed under reduced pressure. The residue was extracted
with ethyl acetate, washed with KHCO3 solution (5%) and finally with
water, The 13-monoesters 3 obtained were purified by TLC (3/I ether/
petroleum ether). Yields, Rf values and mass spectral data of all 13-monoesters 3 prepared are compiled in Table!. For UV, IR and NMR data compare the 13-monoacetate (above) as representative.

12-deoxyphorbol (1) was eluted with ethyl acetate/benzene = 1/4,

yielding 13 g of material.

Biological assays:

Transesterification of the 12-deoxyphorbol ester fraction

To a solution of 12-deoxyphorbol ester fraction (15 g) in methanol (50
ml), 10-' M sodium methoxide/methanol (600 ml) was added. After 20

Irritant activity on the mouse ear and tumor promoting activity on the
back skin of mice were determined according to the standard procedures (5, 8). Experimental details for the latter activity may be taken from
Table II.

h, the reaction was quenched with acetic acid and the solvent was
removed under reduced pressure. The residue was chromatographed
on a polyamide (100 g) column using successively 1/3 water/methanol
(elution of crude 12-deoxyphorbol, 1) and 1/1 water/methanol (elution
of residual 12-deoxyphorbol-13-angelate, that was recycled).
12-Deoxyphorbol-13,20-diacetate (2)
To a solution of crude 1(2 g) in pyridine (6 ml), acetic acid anhydnde (3
ml) was added and the mixture was left for 24 h. After work-up, the
product was purified by TLC (1/4 ethyl acetate/benzene); yield: 170

mg. MS: rn/c = 432 (M). UV, IR and NMR data are fully identical
with those published earlier (7).

12-Deoxyphorbol (1) by alkaline transesterification of its 13,20-diacetate (2)

Sodium methoxide in methanol (1 ml; 101 M) was added to a solution

of the diacetate 2(0.5 mmole) in 0.5 ml of methanol. After 15 h, the reaction mixture was neutralized with acetic acid, methanol was removed

by rotary evaporation and the residue was extracted with butanol.

Compound 1, thus obtained, was purified by TLC (1/20 CH3OHI
CH2C12); yield: 90%. MS: rn/c = 348 (M). 'H-NMR (d5-pyridine): 1H: 7.8; 7-H: 6.18 (d, J = 4.5 Hz); 20-H2: 4.3 (s); 8-H, 10.H: 3.7 (m); 5H2: 3.1 (s); 19-H3: 1.7 (m); OH (exchangeable with D20): 5.8 ppm. IR

(KBr): 3400 (OH); 1690 (CO); 1620 cm' (C = C). UV: (methanol)
Xlna (a) = 233 (4600), 332 nm (60).

Acylation of! to 13,20-diesters 2

To a solution of 1(1.5 mmole) in benzene (50 ml), pyridine (2 ml) was
added. A solution of freshly distilled acid chloride (6 mmole) in benzene (6 ml) was gradually added while stirring. After 12 h, the mixture
was worked up. The 13,20-diesters 2, thus obtained, were purified by
TLC (1/3 ether/petroleum ether) except the 13,20-dibenzoate which
was obtained as crystals (colourless needles from acetone, rn. p. 122

Results
12-Deoxyphorbol (1) may be prepared in quantities by isolation from Euphorbia resinifera resin as its 13,20-diacetate (2).
The hydrophobic triterpenes present in the methanol extract of
the resin were largely removed by partitioning between watermethanol and petroleum ether. Removal of the strong hydrophilic substances were affected by filtration on silica gel using
ethyl acetate as mobile phase. Subsequent chromatography on
silica gel lead to the isolation of a 12-deoxyphorbol ester fraction which amounts to 1.3 % by weight of the resin. Alkaline
(0.1 M sodium methoxide/methanol) transesterification of this
fraction provided 12-deoxyphorbol (1, for structural formulas
see Fig. 1) and 12-deoxyphorbol-13-angelate which evidently

resisted transesterification (at least partly). Both reaction

products were separated by column chromatography on polyamide. The angelate was recycled whereas the crude 1 was
converted to its 13,20-diacetate 2, that is more stable than land
hence more suitable for storage.
Acylation of 1 with acid chlorideslpyridine afforded 13,20diesters 2 (Table I). By partial transesterification of the diesters
2 the 13-monoesters 3 were obtained (Table I). The mass spectra of the 13,20-diesters 2 and of the 13-monoesters 3 show the
expected molecular ions (Table I) and a similar fragmentation
pattern. The common ions at m/e = 330, 312 and 294 are characteristic for the 12-deoxyphorbol (1) moiety. The NMR spectra of the esters exhibit signals typical of this class of compounds (7).

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from Merck. For analytical and preparative TLC, Merck silica gel
HF4 and PF4 were used, respectively. For column filtration or chro-

Structure Activity Relations of Polyfunctional Diterpenes of the Tigliane Type, VI

67

Table I. Yields, R, values and molecular ions of the 13,20-diesters 2 and 13-monoesters 3 prepared from 12-deoxyphorbol (1)

13,20-diesters 2

Acid moieties in
2 or3

Yield

Acetic
Hexanoic
Decanoic
Tetradecanoic
Octadecanoic
Docosanoic
Oleic
Elaidic
Linoleic
Benzoic

(%)

80

0.12

32
50
45
30
28
37
35
35
45

0.41
0.53

13-monoesters 3

Molecular ion
(m/e)

Yield
(%)

432
544
656
768
880
992
876
876
872
556

60
45
42
40
42
27
47
45
38
30

0.03
0.14
0.20
0.25
0.30
0.34

0.54
0.64
0.60
0.63

0.28

0.64
0.62
0.37

0.28
0.25

R'

Molecular ion
(m/e)

0.08
0.17

0.32
0.45

0.21

0.51

0.23
0.24
0.28
0.25
0.25
0.24
0.13

0.52
0.53
0.60
0.57
0.57
0.56
0.38

390

446
502
558
614
670
612
612
610
452

a A: ethyl acetate/benzene = 1/4, B: ether/petroleum ether = 1/2

b

A: ether/petroleum ether = 5/1, B: acetone/petroleum ether =

1/1

Table II. Tumor-promoting activities of the 12-deoxyphorbol-13-esters (3) prepared in the standardized assay (5, 8) on the back skin of mice.
Positive control: 12-O-tetradecanoylphorbol- 13-acetate (TPA)

ester(3)

Application
Single
dose p
(nmole)

Acetate
Hexanoate
Decanoate
Tetradecanoate
Octadecanoate
Docosanoate
Oleate
Elaidinoate
Linoleate
Benzoate

TPA

Duration

At 12 weeks

(weeks)

200
40
40
80
100
80
80
80
80
160

45
48
48
42
39
26
23
23
24
48

48

Tumor
ratea

Tumor

0/28
0/28
8/28

0/28

yieldt'

2 or3
Acetic
Hexanoic
Decanoic
Tetradecanoic
Octadecanoic
Docosanoic
Oleic
Elaidic
Linoleic
Benzoic

TPA

13,20-diesters2
(nmole/ear)

0/28
0/24

0/28
0/24

100
86

12/27
17/24
16/27

29/27

96

65/24
51/26
0/24
67/24
77/26
72/27

86

10/27
10/27

25/27
21/27

4/28
0/27

8/28
0/27

22/26
16/27
0/25

8/28

14/28

11/26

0/24
17/24

Table HI. Irritant doses 50 (ID50, according to the standard procedure (5, 8)) of the 13,20-diesters 2 and 13-monoesters 3 of 12-deoxyphorbol (1) prepared as compared to 12-O-tetradecanoylphorbol-13acetate (TPA) (5)
Acid moieties in

Tumor

8/28
0/27

a number of tumor bearing mice/number of survivors.

b total number of tumors/number of survivors.

ID 13-monoesters 3

Survival Tumors invest. / Malig. tumors

intotal"

Tumor
rate

0/28
9/29
28/27
15/28
0/27

11/27

Histological diagnosisc

Promoting activity
At 20 weeks
yield

rate(%) miceinvest.

0/25

96
86
86
93
96
89

25/26

93

3/3
37/17
85/22
79/22

1 PEC
2PEC, 1 ADBAM
1 PEC

70/16
59/16
81/19

1 PEC

0
0

according to the standard procedure (8).

PEC: Squamous Cell Carcinoma, ADBAM: Basaloma

Both 13,20-diesters 2 and 13-monoesters 3 were assayed for

their irritant activities according to the standard procedure on
the mouse ear (5,8). The results are given in Table III. The 13monoesters 3 were further assayed for their tumor promoting
potency according to the standard procedure on the back skin
of mice (5, 8). The results are given in Figs. 2 and 3.

(nmole/ear)
2.3
0.19
0.26
0.050
0.14
2.5
0.087
0.078
0.062
4.4

1.8
1.0

5.4
0.26
5.1
4.0

0.4
23
57

0.016

a standard deviation a: 1.3, significance level a = 0.05

b
not determined

Discussion
Table III shows that the 13,20-diesters 2 generally have lower
irritant activities than the corresponding 13-monoesters 3, except in case of the acetates which are comparably active. The
present set of esters clearly demonstrates that for maximal irritant activity the primary hydroxyl 20 must not be acylated (at
least not with an acyl residue identical with that in position 13).
A very similar relationship was discovered previously when
comparing phorbol-12,13-diesters to the corresponding phorbol-12,13,20-triesters (5, 6, 810). Within the 13-monoesters 3
it can be seen that the length of the acyl chain strongly affects
the irritant activity as is the case with phorbol-12,13-diesters
[bearing identical acyl chains (11)] or with ingenol-3-esters

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12-Deoxyphorbol-13-

68

Planta Medica 1984

Fig. 2 Tumor rates and tumor yields as obtained in the standardized assay for tumor

TUMOR H
YIELD

promoting activity on the back skin of NMRImice (5,8); initiation: once a single dose of i =
100 nmole DMBA; promotion: twice weekly
a single dose p of 12-deoxyphorbol-13-ester
(3), or 12-O-tetradecanoylphorbol-13-acetate

30

1
50

WEEKS

(TPA). 12-deoxyphorbol-13-decanoate: p =
40 nmole; 12-deoxyphorbol-13-tetradecanoate: p = 80 nmole; 12-deoxyphorbol-13-octadecanoate: p = 100 nmole; TPA: p = 5 nmole.

\i
* 13-o4W.
A 13-.SIdMt

Ia-probe.

(%J

Fig. 3 See also legend of Fig. 2. Promotion:

twice weekly a single dose p of 12-deoxyphorbol-13-ester (3). 12-deoxyphorbol-13-oleate:
p = 80 nmole; 12-deoxyphorbol-13-elaidate:
p = 80 nmole; 12-deoxyphorbol-13-linoleate:
p = 80 nmole; 12-deoxyphorbol-13-octadecanoate: p = 100 nmole.

Downloaded by: NYU. Copyrighted material.

40 WEENS

Studies on 0-Methylfiavinantine
Acknowledgements

For histological diagnosis we are gratefully indebted to Prof. Dr. K.

Goerttler, Institut fr Experimentelle Pathologic, Deutsches Krebsforschungszentrum, Heidelberg. The authors wish also to express their
thanks and gratitude to the Alexander von Humboldt Foundation for
the fellowship granted to Prof. Dr. Salah Zayed.

References
(1) Hecker, E. (1977) Pure Appl. Chem. 49, 1423.
(2) Evans, F. J., Soper, C. J. (1978) J. Nat. Prod. 41, 193.
(3) Gschwendt, M., Hecker, E. (1974) Z. Krebsforsch. 81, 193.
(4) Hergenhahn, M., Kusumoto, S., Hecker, E. (1974) Expenentia
30, 1438.
(5) Hecker, E., Schmidt, R. (1974) Progr. Chem. Org. Natur. Prod.
31, 377.
(6) Hecker, E. (1978) in Mechanisms of Tumor Promotion and Cocarcinogenesis, (Slaga, T. J., Sivak, A., Boutwell, R. K., eds.),
Vol. 2, p. 11, Raven Press, New York.
(7) Gschwendt, M., Hecker, E. (1969) Tetrahedron Lett. 3509.

(8) Hecker, E. (1971) in Methods in Cancer Research, Vol. 6,

(Busch, H. ed), p. 439, Academic Press, New York, London.
(9) Schmidt, R., Hecker, E., 3rd Mtg. Eur. Assoc. Cancer Res.,Nottingham 23.9.25.9. 1975 (Abstr., p.63).

(10) Hecker, E. (1968) Planta Med., 24, (Suppl.).

(11) Thielmann, H. W., Hecker, E. (1969) in Fortschritte der Krebsforschung, Bericht ber die 10. wissenschaftliche Tagung des
Deutschen Zentralausschusses fr Krebsbekampfung und Krebsforschunge. V., (Schmidt, C. G., Wetter, 0., eds.), Berlin 1968,
p. 171, Schattauer Verlag, Stuttgart, New York.
(12) Sorg, B., Hecker, E. (1982) Z. Naturforsch. 37b, 748.
(13) Schairer, H. U. (1966) Dissertation, University of Heidelberg.

Studies on 0-Methylfiavinantine
1. Analgesic Effect of 0-Methylfiavinantine in Mice

R. Ansa-Asamoah"3, G. A. Starmer2

Abstract: The antinociceptive effects of 0-methylfiavinantine (OMF),

a morphinandienone alkaloid, were investigated in the mouse hot plate
and abdominal constriction tests (nociceptive agents: 5-Hydroxytryptamine, acetylcholine, bradykinin, prostaglandin, E1 (PGE1), formic
acid and phenyiquinone).
The potency of OMF in the hot plate test was approximately 10 times less than that of morphine and the effect was naloxone reversible.
In the abdominal constriction test, morphine was 78650 times more
potent than OMF, depending on the nociceptive agent used, but a higher dose of naloxone was necessary to reverse the response to formic
acid.
Pretreatment of mice with reserpine (1 mg/kg, s.c., 24 h) reduced
and a-methyl-p-tyrosine (200 mg/kg, i.p., 3 h) potentiated the antinociceptive effects of both morphine and OMF in the hot plate test.
The results are considered to indicate that OMF possesses centrallymediated antinociceptive activity which is similar to that of morphine.

Introduction
0-methylfiavinantine (OMF) (Fig. 1) is the major alkaloid of
the root bark of Rhigiocarya racemifera (Menispermaceae)
(33). The alkaloid possesses the morphinandienone structure
and its synthesis has been reported (19, 31). Morphinandienone-containing plants have widespread medicinal and recreational uses. The juice from the leaves of Rhigiocarya racemifera is

used in Ghana as a nasal decongestant and decoctions of the

root, bark and seeds are used as an aphrodisiac (18).
NCH3

H3CO
2

Faculty of Pharmacy, University of Science and Technology, Kumasi

Department of Pharmacology, University of Sydney, Australia
Correspondence address: R. Ansa-Asamoah, Faculty of Pharmacy,
University of Science and Technology, Kumasi, Ghana

OCH3

OMETHVLFI.AVINANTINE
Fig. 1.

NCH3

HO

OH
MORPHINE

Downloaded by: NYU. Copyrighted material.

(12). Thus, the 13-tetradecanoate exhibits maximum potency,

comparable to that of 12-O-tetradecanoylphorbol-13-acetate
(TPA, (5)). The 13-oleate, -elaidate and the -linoleate have similar irritant activities as the 13-octadecanoate thus indicating
that in this series non-conjugated unsaturation ot the acyl chain
has no great influence. Analogously to the phorbol-12,13-dibenzoate (6, 13) the 12-deoxyphorbol-13-benzoate shows only
weak irritant activity when compared to a saturated aliphatic
ester with a similar number of C atoms.
Concerning the tumor-promoting activities of the 13-monoesters 3 synthetized (Figs. 2 and 3) the 12-deoxyphorbol-13acetate and the benzoate are not active in the doses tested (Table II). Comparing the hexanoate and the decanoate (both tested with p = 40 nmole) only the latter exhibits tumor promoting activity (Fig. 2). In the doses chosen the tetradecanoate (p
= 80 nmole) and the octadecanoate (p = 100 nmole) seem to be
equipotent and also equipotent with respect to TPA (with p =5
nmole). It should be mentioned that previously (3) the 13-tetradecanoate (with p = 12.5 nmole) was found to be comparably
potent as TPA (with p 10 nmole). From the data taken for
mice of the same strain of the two studies (this and bc. cit. 3)
it might appear that 12-deoxyphorbol-13-tetradecanoate possibly does not show a pronounced dose response relationship. To
check the validity of this impression extended additional studies would be required.
In contrast to the tetra- and octadecanoate, the docosanoate
has no tumor-promoting activity at all, thus indicating that there exists an optimum chain length of the acyl residue. In case of
the oleate, the elaidate and the linoleate (all with p = 80 nmole)
as compared to the octadecanoate (p = 100 nmole, Fig. 3) unsaturation of the acyl chains does not cause dramatic effects.

69