CS277471B6 - A cyclosporin derivatives - Google Patents

Abstract

The present invention relates to novel cyclosporin A derivatives with the immunosuppressive effect they have a functional group that can be attached cyclosporine backbone. These are derivatives formulas: rQ-o6-Abu-Sar-MeLeu-Val-MeLeuAla- (D) -Ala-MeLeu-MeLeu-MeVali where Q stands for either (2S, 3R, 4R) -3-hydroxy-4,7-dimethyl-2-methylamino-8-oxo-octanoic acid acid, or (2S, 3R, 4R) -3-Hydroxy-4-methyl-2-methylamine o-9-oxo-nonanoic acid.

Description

The invention relates to cyclosporin A derivatives with an immunosuppressive effect, in the structure of which an aldehyde functional group is introduced on the first amino acid of the cyclic skeleton of cyclosporin.

Cyclosporin A is a cyclic undecapeptide whose structure is described by the formula: ...

r MeBmt-a-Abu-Sar-MeLeu-Val-MeLeu-Ala- (D) -Ala-MeLeu-MeLeu-MeVal-i,

2 3 4 5 6 7 8 9 10 11 wherein the MeBmt residue is HOOC-CH (NHCH) -CH (OH) -CH _(CH?) -CH, -CH = CH-CH ₇ 12 3 4 5 678 i.e. (2S , 3R, 4R, 6E) -3-hydroxy-4-methyl-2-methylamino-6-octenoic acid.

In addition to cyclosporin A, a number of B to Z cyclosporins have been isolated, which are derived from cyclosporin A by substitution of individual amino acids in the cycle (Helvetica Chimica Acta 70, 13, 1987). These cyclosporins are a natural metabolite of some species of deuteromycetes and are industrially produced by culturing high-production strains, for example Tolypocladium inflatum NRRL 8044 (Eur. J. Appl. Microbiol. Biotechnol. 14, 237, 1982), Tolypocladium terricola CCEB 2-201 M, (MS. U.S. Patent Nos. 2624 and 2625), or Cylindrocarpon lucidum NRRL 5760 (U.S. Patent No. 4,117,118). An alternative, but more demanding, method of preparation is the total synthesis, by means of which some other cyclosporin derivatives have also been prepared (European Patent Nos. 34567 and 194972).

Cyclosporin A is used in human medicine as an immunosuppressant in transplants, in addition, cyclosporins have antiarrhythmic, antiparasitic and aid in the treatment of autoimmune diseases. In addition to therapeutic effects, cyclosporins also show adverse side effects, in particular nephrotoxicity (e.g. Kidney Dis. 1986, 383, 423), which could be eliminated by targeted targeting of drugs consisting of a selective macromolecular carrier and bound cyclosporin. However, the disadvantage of cyclosporins A to Z is that they do not have an active group that could be used for such a bond to the support, except for the -OH group, whose chemical bonding, however, leads to a loss of derivative activity.

The elimination of this disadvantage and the possibility of attaching cyclosporin to the carrier is achieved by a structural modification consisting in the introduction of an aldehyde group into the structure of cyclosporin.

The present invention relates to cyclosporin A derivatives of the general formula:

- Q-a-Abu-Sar-MeLeu-Val-MeLeu-Ala- (D) -Ala-MeLeu-MeLeu-MeVal η, 1 2 3 4 5 6 7 8 9 '10 11 where Q is either

HOOC-CH (NHCH) -CH (OH) -CH (CHO) _{9 -CH-CH?} -CH _(CH?) -CHO, i.e. (2S, 3R, 4R) -3-hydroxy-4,7-dimethyl-2-methylamino-8-oxo-octanoic acid (hereinafter Q] _), or HOOC- CH (NHCH ₃ ) -CH (OH) -CH (CH ₃ ) -CH ₂ -CH ₂ -CH ₂ -CH ₂ -CHO, 1 2 3 4 56789 i.e. (2S, 3R, 4R) -3-hydroxy- 4-methyl-2-methylamino-9-oxo-nonanoic acid (hereinafter only Q ₂ ) ·

Said derivatives have the general formula ^C 63 ^H H3 ° 13 'are positional isomers.

The advantage of these derivatives is that the aldehyde group is the reactive center of the resulting cyclosporin A derivative, which allows the cyclosporin to be attached to the support based on generally known selective reactions of this group, such as condensations, with amines, hydrazines or semicarbazides.

The identity of the substances according to the invention is proved by nuclear magnetic resonance spectra. The presence of an aldehyde group on the cyclic skeleton of cyclosporine clearly results from the presence of signals 5 (H) 9.60 ppm (doublet 1.8 Hz), 9.61 ppm (doublet 1.8 Hz)

- R conformation and S atoms C7, 5 (C) 205 ppm (CDC1 ₃₎ cyclosporin A, wherein the first amino acid is Q signals _LZ and presence of 5 (H) 9.74 ppm (t, 1.8 Hz), S ( C) 203 ppm (CDCl ₃ ) for a cyclosporin A derivative, wherein the first amino acid is Q ₂ . Figure 1 shows the 1 H NMR spectrum of a derivative of the above formula, wherein the first amino acid is Q 1. Figure 2 shows the 1 H NMR spectrum of a derivative of the above formula wherein the first amino acid is Q ₂ . Figure 3 shows the ¹³ C NMR spectrum of a derivative of the above formula wherein the first amino acid is Q ₂ . Comparison of the effect of cyclosporin A and cyclosporin A derivatives, where the first amino acid is or Q ₂ .

The effect of cyclosporin A and its aldehyde derivatives was tested on short-term cultures of mouse lymphocytes. A cell suspension was prepared from the spleens of Balb mice and the cells were ^{transferred in an amount of 2 x 10 5} to 96-well panels with 10 μg / ml concanavalin A medium. % by volume of air. After this time, 1 μθΐ of ³ H-thymidine was added all at once, and after another 8 hours of culture, incorporation into DNA was determined. To compare the effect of cyclosporin A with its derivatives of the above formula, where the first amino acid is or Q2, a concentration of 1 μg to 20 ng of cyclosporin per millimeter of culture medium was added to the measurement wells. Both isomeric derivatives of cyclosporin A gave identical results, which are summarized in Tab. 1.

CS 277471 B6 *

Tab. 1: Effect of cyclosporin A and cyclosporin A derivatives on mouse lymphocyte proliferation.

Concentration% inhibition of proliferation cyclosporin A derivative cyc-A (QL. Or Q ₂ )

000 of 9593

800 of 8987

600 of 7168

400 of 2321

200 of 107

Claims (1)

Cyclosporin A derivatives of the general formula: p Qg-Abu-Sar-MeLeu-Val-MeLeu-Ala- (D) ~ Ala ~ MeLeu ~ MeLeu-MeVal -j, where Q is either, (2S, 3R, 4R) -3-hydroxy-4,7- dimethyl-2-methylamino-8-oxo-octanoic acid, or (2S, 3R, 4R) 3 "hydroxy-4" methyl-2-methylamino- -9-oxo-nonanoic acid.