FR100F - New drug endowed in particular with antimitotic activity. - Google Patents

Description

FRENCH REPUBLIC

MINISTRY OF INDUSTRY

INDUSTRIAL PROPERTY SERVICE

2nd ADDITION

THE SPECIAL MEDICINE PATENT

N ° 2.653 M taken on 10/1/63 P.V. n ° 949.222 N ° 100 CAM

International classification: A 61 k // C 07 c

New drug endowed in particular with antimitotic activity. gI ^ QTH ££ k

Company known as: ROUSSEL-UCLAF residing in France (Seine).

2 if M FI 1967

Requested on October 1, 1963, at 15 10m, in Paris. —---—

Issued by decree of December 13, 1965.

(Official Bulletin of Industrial Property [B.S.M.], No. 3 of January 17, 1966.)

Addition n ° 64 CAM.

In its main application, dated January 10, 1963, for a special drug patent, the Applicant Company described and claimed the use as a new drug endowed in particular with antimitotic activity of 4-hydroxymethyl colchicines of formula:

CH20R

HsCO.

H3CO

H3CO

/ - <

/ Ri SR2

0

In a first addition of March 1, 1963, the Applicant Company described a new derivative of 4-hydroxy-methyl colchicine, 4-acetoxymethyl-colchicine of formula:

HsCO.

w rn / V \ _ /

H3CO! y ^ H3CO

OCHs deriving from the previous formula with the following definitions:

Ri represents hydrogen;

R2 represents an acetyl group;

R represents an acetyl group;

and Z represents a methoxy group.

The present addition relates to, as a new medicament, endowed in particular with sta-thmokinetic activity, 4-formyloxymethyl colchicine of formula:

0

CH2-O-C-H

H3CO.

HaCO] 'y- *. HsCO

V \>

och3

for which R = - C

y

0

H

Ri = H;

R2 represents an acetyl group;

Z represents a methoxy group.

packaged for the use in medicinal weight and the preparations containing.

The basic skeleton of the colchicium derivative is numbered here after Ring Index System RRJ 3923, 2nd Edition, 1960.

4-Formyloxymethyl colchicine occurs as a product crystallized in colorless needles, soluble in alcohol, acetone, benzene, and chloroform, sparingly soluble in ether, insoluble in water, acids and dilute aqueous alkalis.

Its melting point, determined on the Kofler block, is F. = 194 ° C.

66 2191 0 73 938 3

Price of the booklet: 2 francs

{100 CAM / 2.653 M] —2

Its rotatory power is / a./o "- - 83 ° àz 2 (c - 0.5%, chloroform).

4-formyloxymethyl colchicine is prepared as was shown in the first addition to the patent application filed in France on December 4, 1962 by the applicant company and intituCH2OH

H3COS

H3ccr t h3co

H 0

I I

-N-C-CHs

OCHs

The preparation of the starting product, 4-hydro-xymethyl colchicine, was described in the special drug patent filed on January 10, 1963 by the Applicant Company and entitled “New drug endowed in particular with stathmo-kinetic activity”.

As indicated in the above-mentioned patent application, the product is endowed with valuable pharmacological properties. In particular, it has a powerful stathmokinetic action.

It can be used for the treatment of myeloid leukemias, skin neoplasias, digestive tumors, and carcinomas in general.

4-Formyloxymethyl colchicine is used orally, transcutaneously, or in local applications.

It can be in the form of injectable solutions or suspensions, packaged in ampoules, in multi-dose vials; solutions or suspensions to be prepared at the time of use from a sterile powder; tablets, coated tablets and ointments.

The useful dosage ranges between 5 and 50 mg per day in adults depending on the route of administration.

The pharmaceutical forms such as injectable solutions or suspensions, sterile powders, tablets, coated tablets and ointments are prepared according to the usual methods.

Pharmacological study of the drug object of the invention: Antimitotic activity.

The antimitotic activity of the drug, object of the invention, was studied according to the method described by H. Jequier, D. Branceni and M. Peterfalvi (Arch. Int. Pharmacodyn. 1955, 103, 243).

This method is based on the following facts:

In the rat, the mitotic index of the femoral bone marrow (cells in mitosis / total number of cells) is approximately the same on smears taken in various regions and on several fields "Derivatives of colchicine and thiocolchicine and process of preparation ".

The principle of the preparation consists in reacting formic acid with 4-hydroxy-methyl colchicine, according to the following scheme:

O

CH2-O-C-H

H8COn

HsCO '

HsCO

\

HO

I II -N-C-CHs

XjX \ Q

OCHs from the same smear; this mitotic index is normally between 10 and 20 ° / 0oî

After subcutaneous injection of an antimitotic colchicine such as, for example, colchicin which blocks the division of cells at an intermediate stage of mitosis, this index rises and reaches its maximum around the sixth hour.

The observed increase depends on the dose administered; the correlation is clearly reflected in an S-shaped curve whose inflexion point is located around the 100 ° / 00 mitotic index.

As a measure of the stathmokinetic activity of a product, the antimitotic dose 100 (DAM 100) defined as being that which provides 100 cells in mitosis per 1000 cells is therefore chosen. The standard technique used is as follows: 1. Subcutaneous injection of several doses c the substance to be studied in groups of 2 or 3 rats, the volume injected always being 0.2 cm3 poi 100 g of body weight;

2 ° Sacrifice of the animals, six hours after the i: jection; removal of femoral marrow, preparation of smear and staining with May Griinwal Giemsa; count of 1000 cells on each preparation;

3 ° Determination of the DAM 100 from a graph representing the mitotic indices found as a function of the logarithm of the dose.

The MAH100 of 4-formyloxymethyl colchicii is 14 mg / kg. Under the same conditions, the antimitotic dose (DAM100) is 0.7 mg / kg for colchicine.

Determination of toxicity:

The toxicity test was carried out on 1 Rockland strain mice, weighing between 18 and 22 g.

4-formyloxymethyl colchicine, put in. if; aqueous pension, was administered intraperitoneally, to groups of mice, in increasing backs.

- 3

The animals were kept under observation for a week.

The lethal dose (LD50) is approximately 250 mg / kg. By way of comparison, the LD50 of colchicine is equal to 2 mg / kg.

The comparison of the active doses (DAM100) and the toxic doses (LD50) of 4-formyloxymethyl colchicine and colchicine shows that the

- [100 CAM / 2.653 M]

compound covered by the patent has a significantly more favorable therapeutic margin than that of colchicine.

ABSTRACT

The object of the present addition is, as a new medicament, endowed in particular with stathmokinetic activity, 4 - formyloxymethyl colchicine of formula:

0

CH2-O-C-H

H3CO

H3CO | At HsCO

HO

--N-C-CHs

\

y \

och3

F = 194 oc.

/ oc / ;, 0 = - 83 ° ± 2 (c = 0.5%, chloroform) packaged for use at medicinal weight.

4-Formyloxymethyl colchicine presented, in particular in the form of injectable solutions or suspensions, packaged in ampoules, in vials

multiple socket; solutions or suspensions to be prepared at the time of use from a sterile powder; tablets, coated tablets and ointments.

Company known as: ROUSSEL-UCLAF

DOCUMENTARY NOTICE ON THE NEW FEATURE

Documents likely to affect the novelty of the medicinal product: none.

Documents illustrating the state of the art in this area:

- Reports from the Académie des Sciences, n ° 22, 255, p. 2983, Colchici-nic series formylation, M.-L. Velluz.

To sell the booklets, contact the Imprimerie Nationale, 27, rue de la Convention, Paris (15th).