DK144420B - METHOD OF ANALOGUE FOR THE PREPARATION OF WINCALCALOID DERIVATIVES AND SALTS THEREOF - Google Patents

Description

(19) DENMARK ijllO

±

Ip (12) PRESENTATION DI) 144420 B

DIRECTORATE FOfl PATENT AND TRADEMARKET SYSTEM

(21) Application No. 224/78 (51) | nt.CI.3 C 07 D 519/04 (22) Submission date ^ 7 · Jan. 1978 (24) Race day 17 · Jan. 1978 (41) Aim. available July 20th. 1978 (44) Submitted Mar 8 1982 (86) International Application No. ~ (86) International Filing Day ~ (85) Continuation Day ~ (62) Stock Application No., -

(30) Priority 19 · Jan. 1977 * 760595, US 25 Nov. 1977, 855979, US

(71) Applicant ELI LILLY AND COMPANY, Indianapolis, US.

(72) Inventor Gerald Lee Thompson, US.

(74) Associate Engineer Hofman-Bang & Boutard.

(54) Analogous process for the preparation of vinca alkaloid derivatives and salts thereof.

The present invention relates to an analogous process for the preparation of vinca alkaloid derivatives having the general formula II as claimed in the preamble of claim 1, and to pharmaceutically acceptable salts thereof, and the process of the invention is characterized by

GQ

equal to the characteristic part of claim 1.

CM

J- -d- -d- *

Q

2 144420

Several naturally occurring alkaloids recoverable from Vinca rosea have been found effective in the treatment of experimentally induced malignant conditions in animals.

Among these, leurosine (U.S. Patent No. 3,370,057)> vincaleukoblastin (vinblastine), hereinafter referred to as VLB (U.S. Patent No. 3,097,137), leurosidine (vinrosidine), and leurocristine (VCR or vineristin) are both in U.S.A. U.S. Patent No. 3,205,220), 4, -desoxy-VLB "A" and "B", Tetrahedron Letters, 783 (1968) (desacetyl-leurosine hydrazide is also mentioned herein); 4-desacetoxy-vinblastine (U.S. Patent No. 3,954,773); 4-desacetoxy-3'-hydroxyvinblastine (U.S. Patent No. 3,944,554); leurocombin (U.S. Patent No. 3,890,325)> leuroformin (N-formylleurosine; see Belgian Patent No. 811,110) and vincadiolin (U.S. Patent No. 3,887,555). Two of these alkaloids, VLB and leurocristine, are now marketed as drugs for the treatment of malignant conditions in humans, especially the leukemias and related diseases.

The dimeric alkaloids prepared from Vinca rosea can be depicted by the general formula I: 5 'R3, R3- R4 Formula 1 \? 5 V \ -c-0-ch3 14 i; ii h; o! 9 / * \! I-f in IA Vv '·· ™ / "3> 2:

R 6-0-CH3 II

In Formula I wherein R is acetoxy, R is methyl, R 1 is hydroxyl, 4 1 R is ethyl and R is H, VLB is depicted; if R is acetoxy, R 3 is formyl, R is hydroxyl, R is ethyl and R 4 is H, vincristine is depicted; if R is acetoxy, R is methyl, R 1 is ethyl, R 5 is hydroxyl, and R is H, leurosidine is depicted; if R is acetoxy, R 1 is methyl, R 3 and R 3 are H and R 2 is ethyl, 4, -desoxy-VLB- "A" is depicted; if R, R and R are the same as in 4'-deoxy-VLB-MA ", but R 1 is ethyl and R 2 is hydrogen, 4, -desoxy-VLB-" B "is depicted; 5

and if R is acetoxy, R is methyl, * R 1 is ethyl, and R and R

taken together forming an α-epoxide ring, leurosine is depicted.

Of the above alkaloids, vincristine is the most useful and at the same time the least recoverable from Vinca. Jovanovics et al., U.S.A. Patent 3,899,493 'recently developed an oxidative method for converting the relatively more abundant VLB to vincristine by chromic acid oxidation at low temperatures (-60 ° C). In the vinca alkaloid fraction containing dimeric indole dihydroindoles, there are other relatively abundant alkaloids such as leurosine, and it would be desirable to convert these directly or indirectly to vincristine or to a drug having similar tumor-degrading effect. It is well known that leurosine can be converted to 4'-deoxy-VLB "B" (together with alternating amounts of 4, -deoxy-VLB - "A") by treatment with Raney nickel under reflux in absolute ethanol, see Neuss, Gorman, Cone, and Huckstep, Tetrahedron Letters 783-787 (1968). While leurosine exhibited tumor-degrading activity against experimentally induced tumors in animals, the clinical response was limited. 4, -desoxy-VLB- "A" and 4, -deoxy-VLB- "B" were reported as having no reproducible effect against experimentally induced tumors in mice.

It has now been found that, in the process according to the invention, derivatives of 4 * -desoxy-VLB- "A" and "B" obtained from 4, -desoxy-VLB- "A" and "B", -desoxy-VLB- "A" and "B" themselves have tumor-degrading effect, and thus can be used as drugs, see below.

A compound of the general formula I wherein R 1 is ethyl, R 2 is acetoxy, R 2 is CHO, and R 1 'and R 2 are hydrogen are designated as 4'-deoxyvinocristine; a compound wherein R R is hydroxy, while the other groups are the same, is designated as 4, -desoxy-4-desacetylvincristine.

4 UU2 €

Since it is not known to the vineristin-like alkaloid, which has the opposite configuration of hydrogen and ethyl at the 4'-34 carbon atom, such compounds wherein ϊν is ethyl and R is hydrogen are referred to with reference to the compound leurosidine. having the same configuration at the 4'-carbon atom as 4, -desoxy-VLB- "Bn, and they will be referred to as derivatives of 1-formylleurosidine; for example, 4, -desoxy-1-formylleurosidine (or 4 '-des-oxyepivineristin) and 4'-deoxy-4-desacetyl-1-formylleurosidine, wherein R 1 is acetoxy and hydroxy, respectively. In each of the names listed above, it should be understood that the 1-methyl group in leurosidine has been replaced by a formyl group and that the term "1-desmethyl" has been omitted to simplify the nomenclature.

Nontoxic acids useful for forming pharmaceutically acceptable acid addition salts of the compounds obtained by the process of the invention include salts derived from non-toxic. ganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydrogen iodide acid, nitric acid, phosphoric acid and the like, as well as salts of non-toxic organic acids including aliphatic mono- and dicarboxylic acids, phenyl substituted alkane carboxylic acids, hydroxy acids, hydroxy acids, hydroxy acids, hydroxy acids Thus, such pharmaceutically acceptable salts include sulfate, pyrosulfate, hydrogen sulfate, sulfite, hydrogen sulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, iodide, iodide, iodide, iodide, iodide, caprylate, aerylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, phthalate, phthalate chlorobenzenesulfonate, xylene sulfonate, phenylac etate, phenylpropionate, phenylbutyrate, citrate, lactate, 2-hydroxybutyrate, glycollate, malate, tartrate, methanesulfonate, propane sulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate and similar salts.

In the chromic acid oxidation of the invention at low temperature in acidic environment, e.g. chromium trioxide and acetic acid are used.

^ 144420

The process according to the invention is further illustrated by the following example.

EXAMPLE

Preparation of 4'-deoxyvinocristine 582 mg of chromium trioxide is dissolved in 5.8 ml of acetic acid and 0.6 ml of water.

This oxidizing mixture is added drop by drop over 5 minutes to a stirred solution of 462 mg of 4'-deoxy-VLB "A" in 58 ml of acetone and 2.9 ml of glacial acetic acid at a temperature of approx. -50 ° C. The reaction mixture is kept under stirring at this temperature for approx. 30 minutes, and then cooled to -65 ° C, at which temperature the reaction is quenched by the addition of 12 ml of 14 N aqueous ammonium hydroxide. The basic reaction mixture is then poured into 400 ml of an ice / water mixture and the aqueous layer is extracted with 150 ml of ether followed by three extractions with each 150 ml of chloroform. The organic layers are combined and the combined organic layers are washed with dilute aqueous sodium hydrogen sulfite, separated and dried. Evaporation of the organic solvents leaves as evaporation residue 4 * -desoxyvincristine. Chromatography of the residue on 50 g of silica "activity I" is used to further purify the desired compound. The chromatogram is developed as follows: 300 ml of 3: 1 ethyl acetate / methanol followed by 300 ml of 1: 1 ethyl acetate / methanol. After an initial fraction of 100 ml, 20 ml fractions are collected. The fractions 8-20 are combined. Evaporation of the solvents from the combined fractions leads to 297 mg of light brown solid, which, by thin layer chromatography, appears as essentially a "single spot material" (pure material).

The free base 41-deoxyvinocristine prepared in this way exhibits the following physical characteristics:

Mass spectrum: m / e 808 (M +), 806, 707 IR spectrum: 3465, 1745, 1687, 1220 cm 3 UV spectrum: 210, 222, 255, 290, 298 nm 100 MHz MMR spectrum: methyl single band at d 3.88, 3.67 and 2.07.

6'-Deoxyvinocristine as a light brown solid is dissolved in acetone and the acetone solution is treated with 0.96 ml of 0.36M (2% v / v) sulfuric acid in absolute ethanol. This results in a green solution which is kept overnight at about 0 ° C. Crystallization is induced by scraping or grafting and the solid crystalline 4 * -desoxyvincristine sulfate is separated by filtration. The filter cake is washed with cold acetone. The sulfate salt is somewhat soluble in acetone so that the filtrate is evaporated to dryness and the resulting evaporation residue is recrystallized from ethanol. The crystalline 4'-deoxyvinocrystine sulfate thus obtained from ethanol is filtered and the filter cake is washed with ethanol. Total yield of 4 * -desoxyvincristine sulfate is 226 mg.

Similarly, 794mg of 4, -desoxy-VLB -, Bn can be oxidized with 900 mg of chromium trioxide in 10 ml of glacial acetic acid and 1 ml of water leading to 4'-deoxy-1-formylleurosidine. Thin layer chromatography of the residue evaporated directly from the oxidation mixture prior to purification shows the presence of a large and a small spot plus traces of other components. Recrystallization of the evaporation residue from anhydrous ethanol largely results in a single stain crystalline material which is isolated by filtration and the crystals are washed with cold ethanol.

Chromatography of the free base thus formed through 50 g of silica gel using a 1: 1 methylene chloride / ethyl acetate solvent system containing 20, 30, 45 and 60% by volume of methanol as the eluent according to the following scheme:

System Quantity 1: 1 20% 200 1: 1 30% 100 1: 1 45% 100 1: 1 60% 400 leads to the following fractions: 7 144420

Fraction Eluate Volume 1 160 ml 2 100 "3 50" 4 50 "5 50" 6 120 "7 120"

Fractions 4-7 are combined, leading to 597 mg of a light brown evaporation residue which then leads to 435 mg white, crystalline 4, -desoxy-1-formylleurosidine (from ethanol). The compound showed the following physical characteristics:

Mass Spectrum: m / e 808 (M +), 806, 777, 775, 336, 138, 136.

IR spectrum: (CHCl3) 3470, 1743, 1690, 1222cm-1.

UV spectrum: (C C OHH)) 210, 222, 254, 290, 298.

100 MHz NMR spectrum: methyl single band at δ 3.87, 3.65 and 2.07. pK * = 9.0 and 4.9 (in 66% DMF).

Cl

The sulfate salt is formed by dissolving 435 mg of the free base in 10 ml of hot ethanol and adding 1.5 ml of 2% sulfuric acid in ethanol.

Crystalline 4, -desoxy-1-formylleurosidine sulfate is precipitated by cooling.

The compounds of the present invention which can be depicted by the above formula II, especially those wherein R 1 is acetoxy, are potent anti-tumor agents. In detecting the action of these agents against mouse transplanted tumors, a method was used which involved administering the agent intraperitoneally at a predetermined dosage level for 7 to 10 days after the inoculation of the tumor, or alternatively on the first, fifth and ninth day after inoculation.

Table 1 shows the results of a number of experiments in which transplanted tumors in mice were successfully treated with a compound of the invention.

In the table, column 1 shows the name of the compound, column 2 shows the transplanted tumor, column 3 the dosage level, or dosage level range, and the number of days in which the dosage was administered; column 4 the method of administration, and column 5 the percent inhibition of tumor growth or the percentage extension of survival time, e.g. for B16. (ROS is an abbreviation for Ridgeway's osteogenic sarcoma; GLS for Gardner Lymphoma sarcoma; P1534 (<J) L1210 is leukemia is; CA755 is an adenocarcinoma; and B1 is a melanoma).

9 14442 0: 'Η cd ω ϋ φ d • η cq ω 3 Η ϋ g 0) Ή æ ω> -µ fig ”I & Ί -a νο · 3« ο ^ · 3 -aa jj ch> -Η < t Ή ΙΟ t \ l Ή VO Ο · Η · Η · Η · Η ri ί Μ Η CQCTi [> - CQ <fnl m CQ CQ 2 φ £ -ι Η, Χ IXIIXII ΟΟΧΧΧ + Ο Χ Ο S h O® < | -IOI> -OIOC0 οσνΙΛ ο Ο ο Ο 2 in 2ί5ιΕ OHO) EH CT \ m 0- <Ι "EH m Η Η- Η Η Εη Η Εη ΕΟ HCTiéh

Eh Η!> Οι 03 Ο • Π Φ> 03 03 0! γΗ 0)> • Η Μ 03 Ρ-ι __________ ηη = = = = = = = = = = = = - - - - - ο Η Ο CFV Η 03 ΟΧ m

bi Η Μ X

cd X m X ,.

ΙΟ Μ ΙΟ 03 Ο CO X „uo tn ΗΟΗΗΌΐη tn no tn m X m ·> m m m ·> · * - m m, νώ ,, m” o X-OOXOO ^ ,, ^ Χ-Χ ,. *! *! ωχιχχ οιι ii XX ο X ^ X m miMDtnoocom oo miin oqin \ cnwvo-iwoiooHOW h <r co w CM_ ^ ^ ^ S ooooooooooo o 'o 6 6 oo ooo H' ^ 03

H ^ h) 'S

S 3w S

S S A ii S &

HOin in a co co coca H

3 H m H H 3 J HQ ^ EHpqt> - 0 (0) 00 {OOO ω o

CQ

I ^

_j I * H

& s f

+3 ί | H

φ £ -P S to g · Η 03 · Η Η mo so 0) Ή β ω · Η φ • ti m u J 5 η I O O HO Φ H h Ή I j4 π

03 I 3 03 H 2 H

mom i φ.

H b H cd H ft 03 X Η Π X H j5

Η Ο> 5 °>? + J H

d m s ή m e cd H

Η Φ E-i £ Φ ft ^ 0

£ 1 Η Ο Ή HOH

Eh I Η I tH 3 H

o - i ocd “It? .

A <t H ft -ί Η I + ^ 144420 10 ø Μ dead • h m <u g ri tj

S 0 H

§ bO -P

Ά d ω æ ø φ η φ • Η d γΗ> Ο Q) -Ρ <Η> + Ρί, Χ, Χ + .X _ d æ η ίο Ο a Ο φ + ίο Ο φ ^ ri -d * Ή Ο * Η Ο d -d "C \ i Η Ο Ο 0 d Η Φ Η Φ Η Φ 0" \ Ό Φ rri

ΟΗΦ 1 Pd I -id 1 Arrow I, Χ I

dHi> OOlOOO Ο W ri O <t in

(¼ 0 O to EH H E-l JO B Φ EH W W

• r: Φ t> Φ Φ Φ

H

Φ ί> • rri b £ τ)

S CU

HH = = = = = = = =.

bO

ø O O Ti rri rri II ΙΟ ΙΟ Φ 0 IO CT \ Or tO Ti bt, X to ro x X d ø X X, X Λ φ tj to x X m o ro

σι od η c \ ivo <t-ro H

free λ λ · »γ <λ fc \ νο * · λ ro ** OHO ~ - Ο Ο, Ο X Μ r-N b £ I 1 1 X Ο Ο I I X I o

-p, Χ O LO Ό I I 03 in IOOO

0 ^ rOCMOCT \ H <t-HHOCMH Φ (Q y «V n ft K tv K #» ♦. ·, Tv -P aooooooooooo d d Φ O CTi

<H

φ Φ

H 'd-P

d m Η Φ

Destiny> -P

, Q O φ Φ cd Svoio cQvocn Hd E-l d ri in (PI rri i — I d B B ffl Iri O (3 O 0Φ

> Oh

o τί Φ ··

I * H bQ

H, -PS

I ί >> H bO H

d -p b d h • H ø S cd Ti • P Od H d

Φ 0 O -P CO

• rri Φ H 0 0, d d 0 I Ή die

O Ti Η H 0, Q

d ii.d h

EH -d "Η Φ Η -P

φ!> i t> i d ø h b b -S r! d, x ø X X -P Ti Ød

Ti O O 0 H HH

die-p φ a φ η φ H Øø 0ΦΟ ød, Q d H Ti 0 d o

d I H I Ti d H (X

o - d - in island (ii -d "Φ -d" Η H ri- ri- ri- 11 144420

By using the present novel compounds as anti-tumor agents, they can be administered either parenterally or orally. Upon oral dosing, a suitable amount of a pharmaceutically acceptable salt of a base of formula II formed with a non-toxic acid, such as the sulfate salt, is mixed with starch or other excipient and the mixture is filled into telescopic gelatin capsules each containing between 7 »5 and 50 mg of active ingredients. Similarly, the anti-neoplastic active salt can be mixed with starch, a binder and a lubricant, and the mixture can be switched to tablets each containing from 7 to 5 mg of salt. The tablets may be provided with partial grooves if smaller or partial dosages are to be used. However, parenteral administration is preferred. For this purpose, isotonic solutions containing 1-10 mg / ml salt of formula II such as the sulfate salt are used. The compounds are administered to an extent of from 0.1 to 1 mg / kg, preferably 0.1 to 1 mg / kg, animal body weight once or twice a week or every other week depending on both the activity of the agent and its toxicity. An alternative method for determining the therapeutic dosage is based on the body surface area, and is administered at doses of between 0.1 and 10 mg / m of animal body surface every 7 or 14 days.

In the clinical application of a compound of the invention, the treating physician will initially administer the compound in the same manner and in the same medium and probably against the same tumor types that have been indicated for vineristin or VLB. The dosage levels used would reflect the difference in dosage level found in the treatment of experimentally induced tumors in mice, the dosage levels of the compounds of the invention being significantly lower than those used with vineristin and VLB. As with other antitumor agents, particular attention will be paid in clinical trials to the oncolytic (tumor-removing) effect of the compounds of the invention against the ten listed "indicator" tumors listed on page 266 of "The Design" of Clinical Trials in Cancer Therapy "edited by Staquet (Futura Publishing Company, 1973).

Claims (3)

12 U4A20 Patent Claims. 1. Analogous Process for Preparing Vinca Alkaloid Derivatives of General Formula II s * 5 li y-C-O-CHs i4 I II Η I 0.! e / \ «Ί! fs ffrr ~ r 5t ..... 092 CHsO-t6 in ice / i-R1 V ° YYoH ^ · CH0 6-0-CH3 II o wherein R ^ is OH or 0-C-CH,; and wherein II> O x 4 one of the groups R and R is H and the other is CgH 2; and pharmaceutically acceptable salts thereof; characterized in that in a low temperature reaction mixture, a vinca alkaloid derivative of the general formula III is reacted with the formula III-V / f VV 5> / X 0H3 14 'i II H j 0 IA 1 9 / s \! Wherein R, or R has the meaning given above, with a chromic acid oxidant, and recover the free rabbit or a pharmaceutically acceptable salt thereof. Process according to claim 1 for the preparation of 4'desoxyvinocrystine, characterized in that 4 * -desoxy-VLB- "A" is reacted with chromium trioxide and acetic acid. Process according to claim 2 for the preparation of 4'-deoxyvin-cristine sulphate, characterized in that 4'-deoxy-VLB - "A'f is reacted with chromium trioxide and acetic acid and the product is recovered as a sulphate salt.