JPS6038378B2 - anticancer drug - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel anti-coronavirus agent, more specifically, a compound represented by the following general formula (1).
[In the formula, ■ represents a neocarzinostatin residue, R. 10R2
The present invention relates to an anticancer agent containing a neocarzinostatin derivative (representing a styrene-maleic acid copolymer residue having a molecular weight of 2,500 to 80,000) as an active ingredient.

Neocarcinostatin is derived from Streptomyces carcinostaticus variant F-41 Croya (Strept.
omyces carzjnos is ticus var
．． It is a proteinaceous anti-inflammatory substance produced in the culture of F-41Kuroya (Japanese Patent Publication No. 42-21752,
(U.S. Pat. No. 3,334,022), whose primary structure was determined by one of the inventors, Maeda, and has a total number of amino acid residues of 10.
9 with an estimated molecular weight of 10,700 (ScieMe, Vol. 178, pp. 875-876, 197
2 years and ~ch. Bi ratio hem. Bioph$. 16
3 Kago, pp. 379-Mada 5).

In the treatment of cancer, metastasis of cancer cells is the most important problem, and especially lymph node metastasis is the biggest problem. Therefore, the present inventor conducted various studies to reduce the toxicity of neocarzinostatin, increase the durability of its drug efficacy, and specifically transfer the drug to the lymphatic system. The present inventors have discovered that a compound of formula (1) obtained by reacting two existing free amino groups with a water-soluble styrene-maleic acid copolymer having a molecular weight of 2,500 to 80,000 solves the above object, and has completed the present invention. .
Therefore, the present invention provides an anticancer agent containing a neocarzinostatin derivative with reduced toxicity and which specifically accumulates in the lymph system.

The neocarzinostatin derivative of the present invention (represented by 10-manjo) is produced by reacting neocarzinostatin with a styrene-maleic acid copolymer containing at least one maleic anhydride residue in its molecule. The styrene maleic acid copolymer has a molecular weight of 2,500 to 80,000.
It is preferable that the degree of polymerization is about 5 to 40 as a maleic acid residue, and one of the 7 to 8 leicic acid residues is a maleic anhydride residue.

The reaction between neocarzinostatin and the styrene-maleic acid copolymer is preferably carried out in neutral or weakly basic conditions at a temperature of 0 to 40°C. In doing so, the two free amino groups of neocarzinostatin react with the maleic anhydride residue moiety of the styrene-maleic acid copolymer (to give a compound of type 1. Regarding the structure of neocarzinostatin, Scieme 17 group lid 8 already published in 1972
75-876, and according to this, the only free a/groups present in the side chains of polypeptides are alanine at the 1st position and lysine at the 20th position of the constituent amino acids. In the above reaction, the two free amino groups of neocarzinostatin react with the maleic anhydride residues of the styrene-maleic acid copolymer to form an acid amide bond to give the compound of formula (1). Next, the biological activity of the neocarzinostatin derivative obtained in Production Example 1 for the H-form with a molecular weight of about 15 and the L-form with a molecular weight of about 15,000 is as follows.

Furthermore, acute toxicity in rats was significantly reduced by about 1'50 for the H-form and about 1/10 for the L-form compared to neocarzinostatin when administered intravenously.

In order to administer the neocarzinostatin derivative of the present invention to humans, administration methods include local tissue administration such as the primary site of cancer, post-surgery natal site, intradermal, subcutaneous, intramuscular, intravenous, oral administration, etc. , and external administration methods such as topical application and suppositories are preferred.

The dosage varies depending on the administration method, the malignancy of the cancer, the type of cancer, the patient's medical condition and general condition, the progression of the cancer, etc., and may be used for the purpose of preventing lymph node metastasis after surgery, etc., or for therapeutic purposes. It varies depending on the situation, but for example, 0.1 to 10 once a day.
It is preferable to administer 9/k9 mainly once or twice a week or every day. In addition, for example, in the case of intravenous injection, the neocarzinostatin derivative is dissolved in physiological saline and administered as an injection solution at 1 dose per day for adults. It is also possible to further increase the dosage for topical application and oral administration. Next, a manufacturing example will be given and explained.

Production Example 1 1.0 g of neocarzinostatin was dissolved in 50 g of a 0.1 M aqueous sodium bicarbonate solution, and a partial hydrolyzate of styrene oleic anhydride copolymer (monomer molecular weight 25
00-5000 mixture) 1 Add a 35% aqueous solution of bamboo little by little and react at 0-40 qC for 60 minutes.

During this time, the pH is maintained at 7.0-9.5. Dialyze the reaction solution using distilled water. The precipitate generated during dialysis with distilled water was removed, and the supernatant was transferred to a Sephadex G-100 column (1.
When the mixture was poured into a solution (5×10 droplets) and eluted with IQhM ammonium carbonate water, two fractions as shown in FIG. 4 were obtained. This is collected, and the high-molecular class is designated as H-form, and the low-molecular class is designated as L-form.
The above precipitate also has the same biological activity as H-form and L-form, and is soluble in dish hM ammonium carbonate. The physical properties of the H-form and L-form thus obtained are as follows.

m Molecular weight H-form Approximately 150,000 L-form Approximately 15,000 ■Melting point H-form 91-94qo (decomposition) L-form 200-22100 (decomposition) [3} Elemental analysis value H-form L-form C 61.69 46.03 C 7.51 7.33 N I. 70 8.18 (4) Solubility (concentration of 2 c/) H-form L-form Neocarzinostatin Ethanol/Stems, Zen'en 10-10 Pyridiso ++ 10-Acetone'5) Amino acid analysis H-form approx. The results of amino acid analysis of approximately 1 female of 10/9, L body after hydrolysis with isohydrochloric acid are as follows.

H-form L-lysine 1 1 Histidine 0 0 Arginine 3 3-Aspartic acid 11 11-Threonine
12 12 serine 1
0 10 Glutamic acid 5 5 Proline 4 4 Glycine 15 15 Alanine
17 171/2 cystine
4 4 balin 1
2 12 methionine 0 0 isoleuicine 1 1 leucine 6 6 tyrosine 1 1 phenylalanine 5 5 tryptophan* 2 2 (Note 1) Numbers are expressed as number of amino acid residues/average molecular weight.

(Note 2) *Determined spectroscopically. (2) Ultraviolet Absorption Spectrum The results of measurement were shown in FIG. 1 by dissolving the L-isomer and the L-isomer in a 0.19M ammonium carbonate solution.

'71 Infrared absorption spectrum measured by the KBd preservative method, the H-form is shown in Figure 2, and the L-form is shown in Figure 3.

■ Conventional method for quantifying free amino groups [R. Fields, MethMinEm”mo
Lo Sen, 25 strokes, 464 pages, 1972, father ademi
cPress, New York] Therefore, neocarzinostatin, L-form and H-form were reacted with trinitrobenzene sulfonic acid in an aqueous solution, and the color tone of the resulting nitrobenzene derivative was quantified spectroscopically to measure free amino groups.

The results are shown in the table below. As mentioned above, since the two free amino groups present in neocarzinostatin have disappeared in the L and H forms, it is clear that the amino groups have reacted with the polystyrene maleic acid copolymer. be.

[Brief explanation of the drawing]

Figure 1 shows the ultraviolet absorption spectra of H-form and L-form;
The figure shows the infrared absorption spectrum of the H-form, Figure 3 shows the infrared absorption spectrum of the L-form, and Figure 4 shows the result when the supernatant obtained by dialyzing the reaction product against water was subjected to Sephadex G-100 column chromatography. shows the elution state of Figure 1 Figure 4 Figure 2 Figure 3

Claims (1)

[Claims] 1 The following general formula ▲ Numerical formulas, chemical formulas, tables, etc. ▼ (In the formula, (N) represents a neocarzinostatin residue, R_
1+R_2 represents a styrene-maleic acid copolymer residue having a molecular weight of 2,500 to 80,000) An anticancer agent containing a neocarzinostatin derivative as an active ingredient.