JPH1095802A - Camptothecine derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain new camptothecine derivs. having an improved antineoplastic effect by combining a comptoecine compd. with a carboxylated polysaccharide through an amino acid or a peptide. SOLUTION: Comptothecine derivs. obtained by combining a compd. of the formula I [wherein two of R<1> to R<5> adjacent to each other form alkylene or each represent H, one of R<1> to R<5> represents -Xn -AlKm -R<6> , other two of R<1> to R<5> each represent H, alkyl or halogen; or two o them adjacent to each other are bonded to each other to form alkylene in which one C is substd. by -Xn -AlKm -R<6> and other three of them represent H, alkyl or halogen (one or two methylenes of the alkylenes may be substd. by -O-, -Sor -NH-; X represents -O- or -NH-; alk represents alkylene; and R<6> represents -NH2 , formula II, -OH or the like)] with a carboxylated polysaccharide through an amino acid or a peptide.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a novel camptothecin derivative exhibiting enhanced antitumor activity. More specifically, the present invention relates to a novel camptothecin derivative obtained by binding a camptothecin compound having an aminoalkoxy group or a hydroxyalkoxy group to a polysaccharide having a carboxyl group via an amino acid or a peptide, and a method for producing the same. The camptothecin derivative of the present invention is delivered to a target action site in a large amount and selectively, and can exert a desired pharmacological action at the site.The antitumor effect of the camptothecin compound is significantly enhanced. In addition, its side effects are reduced, and it is extremely useful as a pharmaceutical.

[0002]

BACKGROUND OF THE INVENTION Camptothecin is a plant alkaloid and has the following chemical structural formula:

Embedded image Irinotecan hydrochloride, which is known to have anti-leukemic and anti-tumor effects
(CPT-11) is already commercially available. However, while this compound has a strong antitumor activity in clinical practice, it has strong side effects like other antitumor agents and the like, and its use is restricted [Cancer and Chemotherapy, Vol.
9 (1994)]. It has also been reported that various camptothecin compounds have been synthesized and have an antitumor activity (Japanese Patent Application Laid-Open Nos. 1-279891 and 5-2).
22048, JP-A-6-87746, JP-A-6-2
No. 28141, Tokiohei 4-503505, Tokuyohei 4-
No. 502017).

[0003] On the other hand, in order to increase the antitumor activity and suppress the side effects of such kind of drugs having strong side effects as much as possible, recently, a so-called drug delivery system for delivering a necessary amount of the drug to a necessary tissue has been proposed.
(Drug Delivery System) technology is being studied.
In particular, in cancer chemotherapy, there is a problem that a sufficient difference in anticancer drug sensitivity cannot be obtained between tumor cells and normal cells, and a targeted drug delivery system for selectively delivering an anticancer drug to a cancer lesion. Research has been actively conducted, for example, doxorubicin-polysaccharide complex (WO 94/19376), doxorubicin-encapsulated liposome [enhancement of effect of anticancer agent and targeting therapy (published by Science Forum Co., Ltd.), p. 227 (198)
7 years)], dextran-conjugated mitomycin [enhancement of anticancer drug effect and targeting therapy (published by Science Forum Co., Ltd.), p. 278 (1987)] and the like.

[0004]

As described above, the camptothecin compound has an excellent antitumor effect and is extremely useful as a medicament, but has the problem that its use is severely restricted due to its strong side effects. is there. Therefore,
There is a demand for the development of a new derivative that retains its excellent drug activity and suppresses the appearance of undesirable side effects.

[0005]

Means for Solving the Problems In order to obtain an excellent camptothecin derivative which eliminates the above-mentioned problems of the camptothecin compound, the present inventors have applied the technology of the drug delivery system described in WO 94/19376. As a result of further study of applying the method described in (1) to a camptothecin compound, a camptothecin derivative in which a saccharide having a carboxyl group is bound to a camptothecin compound having a reactive group via an amino acid or a peptide is desired. The present invention has been found to have the effect described above, and the present invention has been completed.

That is, according to the present invention, the following general formula having an aminoalkoxy group, a hydroxyalkoxy group, etc.

Embedded image [Wherein, R ^{1 to} R ⁵ are (A) two adjacent R ^{1 to} R ⁵ are bonded to each other to form an alkylene group, or both are hydrogen atoms, and the remaining R ¹ out of ^{1 to} R ⁵
One is a -X _n -Alk _m -R ^6, the other two are hydrogen atom,
An alkyl group or a halogen atom, or (B)
Among R ¹ to R ^5, two adjacent but an alkylene group formed by combining with each other, -X _n -Alk _m -R ⁶ are substituted to any carbon atom of the alkylene group, R ¹ ~ the remaining three are hydrogen atoms R ^5, represents an alkyl group or a halogen atom, (a) and one or two methylene groups in the alkylene group in (B) is -O -, - S- or - X may be replaced with NH-, X is -O- or -NH-, Alk is an alkylene group, R ⁶
Is -NH ₂ ,

Embedded image Or m and n are both 0 or 1, or m is 1 and n is 0] to a saccharide having a carboxyl group via an amino acid or a peptide. The camptothecin derivative was found to have remarkably strong antitumor activity and low toxicity.

An object of the present invention is to provide a novel camptothecin derivative comprising a camptothecin compound represented by the formula [I] and a polysaccharide having a carboxyl group bonded via an amino acid or a peptide. Another object of the present invention is to provide a method for producing the camptothecin derivative.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The camptothecin derivative of the present invention includes a camptothecin compound [I] and a polysaccharide having a carboxyl group bonded via an amino acid or a peptide. As a specific example, a part or all of the carboxyl group of the polysaccharide and the amino group of the amino acid or the peptide are in an acid amide bond, and all or a part of the carboxyl group of the amino acid or the peptide and R ^{6 of the} compound [I] and Can be exemplified by a compound having an acid amide bond or an ester bond. More specifically, a part or all of the carboxyl group of the polysaccharide and the N-terminal amino group of the amino acid or the peptide are an acid amide bond, and the amino acid or the amino acid Compounds in which the C-terminal carboxyl group of the peptide and R ⁶ of compound [I] have an acid amide bond or an ester bond can be mentioned.

Each substituent in the compound represented by the formula [I] of the present invention contains the following groups. An alkylene group formed by bonding two adjacent ones out of R ^{1 to} R ⁵ in (A) (even if one or two methylene groups are replaced by -O-, -S- or -NH-); Good) is formed at the 7-position and 9-position, the 9-position and the 10-position, the 10-position and the 11-position, or the 11-position and the 12-position in the general formula [I], and having 2 to 2 carbon atoms.
And six straight-chain or branched-chain alkylene groups (eg, ethylene group, trimethylene group, tetramethylene group, pentamethylene group, hexamethylene group, methylmethylene group, methylethylene group, methyltrimethylene group).

As an alkylene group in which one methylene group of the alkylene group is replaced by -O-, -S- or -NH-, a methylene group at the terminal and one methylene group other than the terminal are -O-, An alkylene group substituted with -S- or -NH- includes, specifically, a compound represented by the formula -O.
-Alk'- (Alk 'means an alkylene group; the same applies hereinafter)
An alkyleneoxy group represented by (for example, a methyleneoxy group, an ethyleneoxy group, a trimethyleneoxy group, a tetramethyleneoxy group, a methylethyleneoxy group);
An alkyleneamino group represented by NH-Alk′- (for example,
A methyleneamino group, an ethyleneamino group, a trimethyleneamino group, a tetramethyleneamino group, a methylethyleneamino group); an alkylenethio group represented by the formula -S-Alk'-
(E.g., a methylenethio group, an ethylenethio group, a trimethylenethio group, a tetramethylenethio group, a methylethylenethio group); an alkyleneoxyalkyl group represented by the formula -Alk'-O-Alk'- (e.g., a methyleneoxymethyl group,
Ethyleneoxymethyl group, trimethyleneoxymethyl group, methylethyleneoxymethyl group);
An alkyleneaminoalkyl group represented by -Alk'- (for example, a methyleneaminomethyl group, an ethyleneaminomethyl group, a trimethyleneaminomethyl group, a methylethyleneaminomethyl group); represented by the formula -Alk'-S-Alk'- And alkylenethioalkyl groups (for example, methylenethiomethyl group, ethylenethiomethyl group, trimethylenethiomethyl group, methylethylenethiomethyl group) and the like.

The alkylene group in which two methylene groups in the alkylene group are replaced by -O-, -S- or -NH- includes a terminal methylene group and a non-terminal methylene group.
Alkylene groups in which two methylene groups are replaced by -O-, -S- or -NH-, and specifically, a compound represented by the formula-
An alkylenedioxy group represented by O-Alk'-O- (for example, a methylenedioxy group, an ethylenedioxy group, a trimethylenedioxy group, a tetramethylenedioxy group, a methylethylenedioxy group); An alkylenediamino group represented by Alk'-NH- (for example, a methylenediamino group, an ethylenediamino group, a trimethylenediamino group, a tetramethylenediamino group, a methylethylenediamino group); represented by the formula -S-Alk'-S- Examples include an alkylenedithio group (for example, a methylenedithio group, an ethylenedithio group, a trimethylenedithio group, a tetramethylenedithio group, and a methylethylenedithio group).

Alk in the group represented by the formula -X _n -Alk _m -R ⁶ is a linear or branched alkylene group having 1 to 6 carbon atoms (for example, methylene group, ethylene group, trimethylene group, tetramethylene group, pentamethylene group, hexamethylene group, methylethylene group, and a methyltrimethylene group), specific examples of the group represented by the -X _n -Alk _m -R ⁶ is, aminoalkyl group (e.g., aminoethyl Oxy group, aminopropyloxy group), piperazinylalkyloxy group (e.g., piperazinylethyloxy group, piperazinylpropyloxy group, piperazinylbutyloxy group, piperazinylpentyloxy group), hydroxyalkyloxy Groups (e.g., hydroxyethyloxy, hydroxypropyloxy, hydroxybutyloxy, hydroxype Tyloxy group), aminoalkylamino group (e.g., aminoethylamino group, aminopropylamino group, aminobutylamino group, aminopentylamino group), piperazinylalkylamino group (e.g., piperazinylethylamino group, piperazinyl group) Propylamino group, piperazinylbutylamino group, piperazinylpentylamino group), hydroxyalkylamino group (e.g., hydroxyethylamino group, hydroxypropylamino group, hydroxybutylamino group, hydroxypentylamino group), aminoalkyl Groups (e.g., aminomethyl group, aminoethyl group, aminopropyl group, aminobutyl group, aminopentyl group), piperazinylalkyl group (e.g., piperazinylmethyl group, piperazinylethyl group,
Piperazinylpropyl group, piperazinylbutyl group, piperazinylpentyl group), hydroxyalkyl group (for example,
A hydroxymethyl group, a hydroxyethyl group, a hydroxypropyl group, a hydroxybutyl group, a hydroxypentyl group), an amino group, a piperazino group, and a hydroxy group.

Further, an alkylene group formed by bonding two adjacent ones of R ^{1 to} R ⁵ in (B) (where one or two methylene groups are —O—, —S— or —NH
- A in may be replaced), the group -X _n -Alk _m -R ⁶ in any of the carbon atoms of the alkylene group is substituted, in the general formula [I], 7-position And 9
9th and 10th, 10th and 11th or 11th and 12th
A linear or branched alkylene group having 2 to 6 carbon atoms (e.g., ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, methylethylene, methyltrimethylene) Wherein one methylene group among the alkylene groups is -O
-, -S- or -NH- substituted alkylene group and two methylene groups are -O-, -S- or -N
Specific examples of the alkylene group substituted with H- include:
In each case, the same as described above can be mentioned. Further, -X _n -Alk _m -R substituted on one of these carbon atoms
^{6 is} also the same as above.

In the above R ^{1 to} R ⁵ , the alkyl group in the remaining groups other than the adjacent alkylene group is a linear or branched alkyl group having 1 to 6 carbon atoms.
(E.g., methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, pentyl group, hexyl group), and as the halogen atom, fluorine, chlorine, bromine And iodine.

Further, as specific compounds, specific examples of the partial structures of the ring A and the ring B in (A) include the following.

Embedded image [Wherein, X, Alk, R ⁶ , m and n are the same as above]

Specific examples of the partial structure of the ring A and the ring B in (B) include the following.

Embedded image [Wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ represent an alkyl group,
X, Alk, R means a halogen atom or a hydrogen atom
⁶ , m and n are the same as described above.] Among them, preferred combinations of R ^{1 to} R ⁵ include the following. (1) R ¹ and R ² are bonded to form a trimethylene group and R ³ is 3-
Aminopropyl group, R ⁴ and R ⁵ is a hydrogen atom (2) R ¹ is piperazinomethyl group, R ² and R ⁵ is a hydrogen atom, R ³ and R ⁴ combine with the ethylenedioxy group (3) R ¹ is An aminomethyl group, R ² and R ⁵ are hydrogen atoms, R ³
And R ⁴ are bonded to form an ethylenedioxy group (4) R ¹ , R ² , R ⁴ and R ⁵ are hydrogen atoms, R ³ is a 3-aminopropyloxy group (5) R ¹ and R ² are bonded Amino-substituted trimethylene group,
R ³ is a methyl group, R ⁴ is a fluorine atom, R ⁵ is a hydrogen atom (6) R ¹ , R ³ , R ⁴ and R ⁵ are hydrogen atoms, and R ² is an amino group

The camptothecin compound according to the present invention
[I] and the “polysaccharide having a carboxyl group” bonded via an amino acid or a peptide are those described in WO 94 /
Including the same materials as disclosed in US Pat.
Polysaccharides that inherently have a carboxyl group in their structure
(E.g., hyaluronic acid, pectic acid, alginic acid, chondroitin, heparin, etc.) and polysaccharides that do not originally have a carboxyl group (e.g., pullulan, dextran, mannan, chitin, mannoglucan, chitosan, etc.) Including those that introduced. Among them, dextran is particularly preferred as the polysaccharide, and its average molecular weight is 20,000 to 400,000, especially 5
It is preferably in the range of 0000 to 150,000 [Gel Permeation Chromatography (GPC) Method, Shinsei Kagaku Jikken Koza Vol. 20, page 7]. In the case where a carboxyl group is introduced into a polysaccharide which does not originally have a carboxyl group, the hydrogen atoms of some or all of the hydroxyl groups of the polysaccharide having no carboxyl group are carboxy-C _1-4 alkyl groups. It means what has been replaced. In addition, the “polysaccharide having a carboxyl group” of the present invention includes a polysaccharide originally having no carboxyl group once treated with a reducing agent, and then a hydrogen atom of some or all of the hydroxyl groups is converted to carboxy-C _1-. Also includes those substituted with ₄ alkyl groups.

The alkyl portion of the carboxy-C _1-4 alkyl group which is substituted for the hydrogen atom of the hydroxyl group of the above polysaccharide may be linear or branched. Carboxy-C
Preferred examples of the _1-4 alkyl group include a carboxymethyl group, a 1-carboxyethyl group, a 3-carboxypropyl group, a 1-methyl-3-carboxypropyl group, a 2-methyl-3-carboxypropyl group, and a 4-carboxy group. A butyl group and the like are mentioned, and a carboxymethyl group and a 1-carboxyethyl group are particularly preferable. In the present invention, it is preferable that the polysaccharide having a carboxyl group is carboxymethylated dextran or pullulan. When the carboxyalkyl group is introduced, the degree of the introduction can be represented by the “degree of substitution” defined as the number of carboxyalkyl groups per sugar residue. That is,

(Equation 1) It can be expressed as. Hereinafter, this degree of substitution is referred to as “degree of carboxymethyl (CM) conversion” when the carboxyalkyl group is a carboxymethyl group. When the polysaccharide is pullulan, dextran or mannoglucan, the degree of substitution is 3 when all the hydroxyl groups are substituted, and the degree of substitution is 0.3.
It is preferably 3 or more and 0.8 or less. When the polysaccharide is chitin, the degree of substitution is 2 when all of the hydroxyl groups are substituted, preferably from 0.3 to 0.8. Unless the polysaccharide originally has a carboxyl group, it is necessary that at least one carboxyalkyl group be present in the polysaccharide molecule. Therefore, compounds having a substitution degree of 0 in this sense are excluded from the polysaccharide of the present invention. Polysaccharides having these carboxyl groups are described in WO 94 /
19376.

The amino acids to be interposed during the binding between the camptothecin compound [I] and the polysaccharide having a carboxyl group include natural amino acids and synthetic amino acids (D-amino acids).
Amino acids, L-amino acids, and mixtures thereof), and may be any of neutral amino acids, basic amino acids, and acidic amino acids. Furthermore, not only α-amino acids, but also β-amino acids, γ-amino acids, ε-amino acids and the like are included. Specific examples include glycine, α-alanine, β-alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, aspartic acid, glutamic acid, lysine, arginine, phenylalanine, tyrosine, histidine, tryptophan, proline, oxyproline. , Γ-aminobutyric acid, ε-
Aminocaproic acid and the like.

In addition to the peptides derived from the above-mentioned amino acids, the peptides include those containing a compound other than an amino acid in a part of the chain. For example, a dicarboxylic acid such as succinic acid, a diamine such as ethylene diamine or a diol such as ethylene glycol may be present in or at the terminal of the peptide chain. In addition, the binding direction of the peptide chain is usually linked from the N-terminus to the carboxyl group of the polysaccharide by an acid amide bond,
When a basic amino acid (e.g., lysine) is present in the peptide chain, the ε-amino group is bonded to the carboxyl group of the polysaccharide, and the α-amino group is bonded to the C-terminus of the peptide chain to form a peptide chain. The coupling direction may be reversed. Such a peptide is one in which two or more amino acids are peptide-bonded, that is, two or more peptide chains, and preferably two to five peptide chains.
Examples of specific peptide chains include, for example, -Gly
-Gly-L or D-Phe-Gly-, -Gly-Gly-,
-Gly-Gly-Gly-, -Gly-Gly-Gly-Gly-,
-Gly-Gly-Gly-Gly-Gly-, -L or DP
He-Gly-, -L or D-Tyr-Gly- or -L or D-Leu-Gly- and peptide chains containing this sequence in the chain (wherein these peptides and peptide chains containing these sequences) Is introduced into the carboxyl group of the polysaccharide). Of these peptides, -Gl
y-Gly-L or D-Phe-Gly-, -Gly-Gly
-, -Gly-Gly-Gly-, -Gly-Gly-Gly-Gly
-, -Gly-Gly-Gly-Gly-Gly-, -L or D
-Phe-Gly and -L or D-Leu-Gly- are more preferred, and -Gly-Gly-L-Phe-Gly is preferred.
-, -Gly-Gly-, -Gly-Gly-Gly-, -Gly-
Gly-Gly-Gly-, -L or D-Phe-Gly- are particularly preferred.

In order to produce the camptothecin derivative of the present invention, a method is generally employed in which an amino acid or a peptide is bound to the compound represented by the formula [I], and then a carboxyl-containing polysaccharide is reacted with the compound. Is done.
By reaction of compound [I] with an amino acid or peptide,
When R ^{6 in the} formula [I] is the formula: —NH ₂ or a piperazino group, an acid amide bond is formed with the C-terminal carboxyl group of the amino acid or peptide, and when R ⁶ is —OH, an ester bond is formed. At this time, it is preferable that other functional groups in the amino acid or peptide not participating in the acid amide bond or the ester bond, such as the N-terminal amino group or other carboxyl group, be protected by a conventional method. Such protecting groups are not particularly limited as long as they are generally used for protecting amino acids.Examples of protecting groups for amino group include t-butoxycarbonyl group, p-methoxybenzyloxycarbonyl group, and carboxyl group. Examples of the group-protecting group include a lower alkyl group (for example, a t-butyl group) and a benzyl group.

The above-mentioned acid amide bond and ester bond can be formed in a conventional manner, for example, in a suitable solvent in the presence of a condensing agent. As the solvent, dimethylformamide, tetrahydrofuran, etc.,
As the condensing agent, dicyclohexylcarbodiimide, 1-
It is preferable to use (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride or the like.

When the amino or carboxyl group is protected on the camptothecin compound to which the amino acid or peptide obtained as described above is bound, if necessary, the protecting group is removed by a conventional method, and then the carboxyl group is removed. The desired camptothecin derivative is produced by reacting a polysaccharide having a group. By this reaction, a part or all of the carboxyl group of the polysaccharide and the N-terminal amino group of the amino acid or peptide bonded to the camptothecin compound [I] form an acid amide bond.

The reaction of a camptothecin compound [I] with an amino acid or peptide bonded thereto and a polysaccharide having a carboxyl group can be carried out according to a conventional method, for example, in a suitable solvent in the presence of a condensing agent. be able to. As the solvent, water, ethanol, dimethylformamide or a mixed solvent selected therefrom, and as the condensing agent, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 2-ethyloxy-1-ethyloxycarbonyl It is preferable to use -1,2-dihydroquinoline or the like.

In the camptothecin derivative of the present invention,
The binding ratio between the polysaccharide and the camptothecin compound [I], which is a medicinal component, is appropriately selected depending on the type of the polysaccharide. In general, the content of the camptothecin compound [I] is preferably set to the following ratio. The polysaccharides are pullulan, dextran,
In the case of chitin, mannoglucan and N-acetyl-de-N-heparin sulfate, 0.1 to 20% by weight is preferable,
Particularly preferred is 2 to 10% by weight. In the present invention, the average molecular weight of the camptothecin derivative (measured by the GPC method) is, for example, 30,000 when the polysaccharide is dextran.
0-500,000, preferably 60,000-200,
000.

The camptothecin derivative obtained by the above method can be converted into a pharmacologically acceptable salt thereof, if desired. Such salts include alkali metal or alkaline earth metal salts such as sodium, potassium and calcium salts, and amino acid salts such as arginine and lysine salts.

The camptothecin compound [I] in the present invention
Include known compounds (for example, see JP-A-1-27989).
No. 1, JP-A-5-222048, JP-A-6-8774
No. 6, JP-A-6-228141, JP-T-Hei 4-5035
No. 05, Japanese Translation of PCT International Publication No. 5-502017), and by a known method, for example, by a method represented by the following reaction formula 1.

Embedded image [Wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ are the same as defined above;
¹ ′, R ² ′, R ³ ′, R ⁴ ′ and R ⁵ ′ are those in which the amino group, piperazino group or hydroxy group in the substituent —X _n —Alk _m —R ⁶ contained in any of them is protected. Except
The same as R ¹ , R ² , R ³ , R ⁴ and R ⁵ respectively]

That is, the aminocarbonyl compound (1) is converted to a known substance, pyranoindolizine (2) (EP-02206).
01-A) and a process known as the Friedlander condensation reaction (Organic Reactions (Organic Reactions)).
ic Reactions) 28 , pp37-202, John Wiley &
Sons. Inc. New York (cf. 1982)), and the protective group is removed to obtain the desired camptothecin compound [I].

In the above-mentioned production method, the group represented by any one of R ¹ ′ to R ⁵ ′: —X _n —Alk _m —R ⁶ , when m is 1 and n is 1, It can be introduced after the Friedlander condensation reaction. That is, instead of the starting compound (1), the corresponding group in the compound (1) is a hydroxyl group.
(HO—) or an amino group (H ₂ N—) is used and subjected to a Friedlander condensation reaction with the compound (2), and the resulting condensation product is added with a compound represented by the formula: R ⁶ ′ -Alk _m −
OH (R ⁶ ′ is a protected amino group, piperazino group or hydroxy group, and Alk and m are the same as those described above) or a reactive derivative thereof (for example, a protected amino group). Alkyl halide, protected hydroxyalkyl halide), and then removing the protecting group gives the desired compound [I].

Starting compound used in the above method
(1) is produced, for example, by the method shown in the following reaction formula 2.

Embedded image Wherein R ¹ ′, R ² ′, R ³ ′, R ⁴ ′ and R ⁵ ′ are as defined above.

That is, the hydroxy compound (a) is treated with an oxidizing agent such as pyridinium dichromate to give a ketone compound (b), which is then catalytically reduced in the presence of a suitable catalyst such as Pd-C. Thereby, compound (1) is obtained.

In the above hydroxy compound (a), -X _n -Alk _m -R ⁶ contained in any of R ¹ ′, R ² ′, R ³ ′, R ⁴ ′ and R ⁵ ′ is m. When 1, n is 1,
Corresponding groups hydroxyl (HO-) or amino group (H ₂ N-)
And a general formula:

Embedded image [Wherein, Alk and R ⁶ are the same as those described above] or a reactive derivative thereof (for example, a substituted alkyl halide).

The camptothecin derivative of the present invention or a pharmacologically acceptable salt thereof has excellent antitumor activity against various tumors, and is especially useful for solid tumors such as lung cancer, uterine cancer, ovarian cancer, breast cancer, Organ cancer (colorectal cancer, gastric cancer, etc.)].

The camptothecin derivative of the present invention or a pharmaceutically acceptable salt thereof is parenterally (eg, intravenously).
It is preferable to administer to a solution (usually, a solution,
Suspension, emulsion).

The dosage of the target substance of the present invention varies depending on the administration method, age of the patient, weight, condition, etc.
Per day, the camptothecin compound [I] (R ⁶ has the formula: —NH
_{When 2} , the hydrochloride, formula:

Embedded image , The hydrochloride or dihydrochloride).
It is preferable to administer in the range of 02 to 50 mg / kg, particularly 0.1 to 10 mg / kg.

[0036]

The compounds of the present invention and the method for producing the same will be more specifically described by the following examples, but the present invention is not limited thereto.

Example 1 Synthesis of camptothecin derivative represented by the following formula

Embedded image [CM-Dextran · Na represents sodium salt of carboxymethyl dextran] (1) Synthesis of 3- (t-butoxycarbonylamino) propanol 6.0 g of 3-aminopropanol was dissolved in 50 ml of methylene chloride and stirred under ice-cooling. Then, 18.3 g of di-t-butyl dicarbonate is added dropwise. After stirring at room temperature for 12 hours, the reaction mixture is concentrated and separated and purified by silica gel column chromatography to obtain 13.98 g of the title compound as a colorless oil.

Yield: 99.9% IR (neat): ν _max ^{cm -1} = 3380, 1790 Mass: m / z = 176 ([M + H] ⁺ ) NMR (300 MHz, CDCl ₃ ): δ ^TMS = 1. 45 (9
H, s), 1.62-1.72 (2H, m), 3.0 (1H, brs),
3.29 (2H, dd, J = 12 Hz and 6 Hz), 3.66 (2
H, dd, J = 12 Hz and 6 Hz), 4.80 (1 H, brs)

(2) 3- (t-butoxycarbonylamino)
Synthesis of propyl tosylate 3- (t-butoxycarbonylamino) propanol 10.
0 g was dissolved in 100 ml of methylene chloride, and 8.66 g of triethylamine and 16.6 g of tosyl chloride were stirred under ice cooling.
3 g are added and the reaction mixture is stirred overnight at room temperature. After concentrating the reaction mixture and dissolving the residue in water-ethyl acetate, the organic layer is separated, washed with saturated saline, and dried over sodium sulfate. After evaporating the solvent, the residue was separated and purified by silica gel column chromatography to give the title compound 1 as a pale yellow oil.
5.37 g are obtained.

Yield: 82% IR (neat): ν _max ^{cm -1} = 3400,3340,1700 Mass: m / z = 352 ([M + Na] ⁺ ) NMR (300 MHz, CDCl ₃ ): δ ^TMS = 1. 42 (9H,
s), 1.78-1.90 (2H, m), 2.45 (3H, s), 3.1
1-3.22 (2H, m), 4.09 (2H, t, J = 6Hz), 4.
5-4.65 (1H, m), 7.36 (2H, d, J = 8Hz), 7.
77-7.83 (2H, m)

(3) 5- [3 '-(t-butoxycarbonyla)
Mino) propyloxy] -1-hydroxy-8-nitro-
Synthesis of 1,2,3,4-tetrahydronaphthalene 1,5-dihydroxy-8-nitro-1,2,3,4-tetra
2.0 g of lahydronaphthalene (J. Med. Chem., 1973,
16(3), 254) was dissolved in 80 ml of dry DMF,
2 equivalents of potassium, 1.4 equivalents of sodium iodide and 3
-(T-butoxycarbonylamino) propyl tosylate
Add 1.4 equivalents. Reaction mixture at 50 ° C. for 24 hours
After stirring, add ethyl acetate, wash with saturated saline, and add sulfuric acid.
Dry over sodium. The residue is purified by silica gel column chromatography.
Separated and purified by chromatography to obtain a pale yellow amorphous powder.
3.05 g of the title compound are obtained.

Yield: 87% IR (neat): ν _max ^cm-1 = 3360, 1695 Mass: m / z = 384 ([M + NH ₄ ] ⁺ ) NMR (300 MHz, CDCl ₃ ): δ ^TMS = 1.36 (9H,
s), 1.57-1.90 (6H, m), 2.52-2.71 (2H,
m), 3.11 (2H, q, J = 6 Hz), 4.07 (2H, t, J = 6
Hz), 5.12-5.17 (2H, m), 6.89 (1H, t, J =
5.5 Hz), 6.96 (1 H, d, J = 9 Hz), 7.68 (1 H,
d, J = 9Hz)

(4) Synthesis of 5- [3 '-(t-butoxycarbonylamino) propyloxy] -8-nitro-1,2,3,4-tetrahydronaphthalen-1-one 5- [3'-( 2.46 g of [t-butoxycarbonylamino) propyloxy] -1-hydroxy-8-nitro-1,2,3,4-tetrahydronaphthalene were dried in methylene chloride 11
Dissolve in 0 ml and remove molecular sieves 3A (molecular s
6.73 g of Hieves 3A) and 1.5 equivalents of pyridinium dichlorochromate are added and heated to reflux. After the reaction,
The reaction mixture was diluted with ether, and the insolubles were removed by filtration through Celite. The filtrate was concentrated and separated and purified by silica gel column chromatography to obtain 1.87 g of the title compound as a colorless powder.

Melting point: 76-77 ° C. Yield: 76% IR (Melt): ν _max ^{cm −1} = 3550,1700 Mass: m / z = 382 ([M + NH ₄ ] ⁺ ) NMR (300 MHz, CDCl ₃ ): δ ^TMS = 1.44 (9H,
s), 2.02-2.20 (4H, m), 2.68-2.73 (2H,
m), 2.92 (2H, t, J = 6 Hz), 3.36 (2H, q, J =
6.5 Hz), 4.12 (2 H, t, J = 6 Hz), 4.78 (1 H, b
rs), 6.95 (1 H, d, J = 9 Hz), 7.39 (1 H, d, J =
9Hz)

(5) Synthesis of 8-amino-5- [3 '-(t-butoxycarbonylamino) propyloxy] -1,2,3,4-tetrahydronaphthalen-1-one 5- [3'-( 3.55 g of [t-butoxycarbonylamino) propyloxy] -8-nitro-1,2,3,4-tetrahydronaphthalen-1-one is dissolved in 160 ml of ethanol, 420 mg of 10% Pd-C is added, and under hydrogen atmosphere, 1.
Stir for 5 hours. The catalyst is removed by filtration, the filtrate is concentrated,
The residue was separated and purified by silica gel column chromatography to obtain 3.56 g of the title compound as a yellow oil.

Melting point: 112-115 ° C. Yield: 83% IR (Nujol): ν _max ^{cm −1} = 3440,3340,170
0.1650 Mass: m / z = 335 ([M + H] ⁺ ) NMR (300 MHz, CDCl ₃ ): δ ^TMS = 1.45 (9H,
s), 1.92-2.67 (4H, m), 2.61 (2H, t, J = 6
Hz), 2.87 (2H, t, J = 6 Hz), 3.35 (2H, q, J =
6.5 Hz), 3.94 (2 H, t, J = 6 Hz), 4.85 (1 H, b
rs), 6.10 (2H, brs), 6.48 (1H, d, J = 9Hz),
6.94 (1H, d, J = 9Hz)

(6) 10- [3 '-(t-butoxycarbonylamino) propyloxy] -7,9-trimethylene- (20
Synthesis of S) -camptothecin 2.03 g of 8-amino-5- [3 '-(t-butoxycarbonylamino) propyloxy] -1,2,3,4-tetrahydronaphthalen-1-one was dissolved in 85 ml of ethanol. , (4S) -7,8-dihydro-4-ethyl-4-hydroxy-1H-pyrano [3,4-f] indolizine-3,6,
800 mg of 10 (4H) -trione and 58 mg of p-toluenesulfonic acid are added, and the mixture is heated under reflux for 17 hours. After completion of the reaction, the solvent was distilled off, and the obtained residue was separated and purified by silica gel column chromatography to obtain 850 mg of the title compound as a pale yellow powder.

Melting point: 225 to 227 ° C. (decomposition) Yield: 50% IR (Nujol): ν _max ^{cm −1} = 3440,3325,175
0,1740,1655,1620 Mass: m / z = 562 ([M + H] ⁺ ) NMR (300 MHz, CDCl ₃ ): δ ^TMS = 1.03 (3H,
t, J = 7.5 Hz), 1.45 (9 H, s), 1.82-2.18
(6H, m), 3.06-3.13 (4H, m), 3.41 (2H, q,
J = 6Hz), 3.79 (1H, s), 4.24 (2H, t, J = 6H
z), 4.9 (1H, br), 5.16 (2H, s), 5.30 (1H, d,
J = 16 Hz), 5.75 (1 H, d, J = 16 Hz), 7.51
(1H, d, J = 9Hz), 7.61 (1H, s), 8.06 (1H, d,
J = 9Hz)

(7) 10- (3'-aminopropyloxy)
Synthesis of -7,9-trimethylene- (20S) -camptothecin hydrochloride 836 mg of 10- [3 '-(t-butoxycarbonylamino) propyloxy] -7,9-trimethylene- (20S) -camptothecin was suspended in 30 ml of dioxane. It became cloudy and 15 ml of 18% hydrochloric acid-dioxane was added dropwise while stirring under ice cooling. The reaction mixture is stirred at room temperature, and after completion of the reaction, isopropyl ether is added and stirred. The precipitated powder is collected by filtration, washed with ether, and dried under reduced pressure. The obtained yellow powder is dissolved in water and freeze-dried to obtain 620 mg of the title compound as a yellow powder.

Melting point: 194 ° C. or more (decomposition) Yield: 84% IR (Nujol): ν _max ^{cm −1} = 1740,1655 Mass: m / z = 462 ([M-Cl] ⁺ ) NMR (300 MHz, d) _6- DMSO): δ ^TMS = 0.88
(3H, t, J = 7.5Hz), 1.81-1.94 (2H, m), 1.
97-2.15 (4H, m), 3.01-3.14 (6H, m), 4.
28 (2H, t, J = 6 Hz), 5.23 (2H, s), 5.43 (2
H, s), 7.28 (1H, s), 7.71 (1H, d, J = 9.5H
z), 7.95-8.08 (3H, brs), 8.03 (1H, d, J =
9.5Hz)

(8) 10- [3 '-(t-butoxycarbonylglycylglycyl-L-phenylalanylglycylamino) propyloxy] -7,9-trimethylene- (20S)
Synthesis of camptothecin 158 mg of 10- (3'-aminopropyloxy) -7,9-trimethylene- (20S) -camptothecin hydrochloride and 49 mg of diisopropylethylamine were dissolved in 5 ml of DMF with stirring, and t-butoxycarbonyl-glycylglycyl was added thereto. -278 mg of L-phenylalanylglycine and N
-143 mg of hydroxysuccinimide in 8 ml of dry DMF
, And further stirred under ice-cooling for 1 hour.
183 mg of-(3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is added, and the mixture is stirred for 16 hours at room temperature. After completion of the reaction, the solvent was distilled off, and the obtained residue was separated and purified by silica gel column chromatography to obtain 285 mg of the title compound as a pale yellow powder.

Yield: quantitative yield IR (Nujol): ν _max ^{cm -1} = 3290,1660 Mass: m / z = 880 ([M + H] ⁺ ) NMR (300 MHz, CDCl ₃ ): δ ^TMS = 1. 02 (3H,
t, J = 7.5 Hz), 1.43 (9 H, s), 1.85-1.94
(2H, m), 2.02-2.10 (4H, m), 2.97-3.05
(5H, m), 3.23 (1H, dd, J = 14Hz and 5Hz),
3.49 (2H, q, J = 6.5 Hz), 3.60-3.80 (6
H, m), 4.20 (1H, t, J = 6Hz), 4.50-4.56
(1H, m), 5.11 (2H, s), 5.29 (1H, d, J = 16.
5 Hz), 5.71 (1 H, d, J = 16.5 Hz), 5.85 (1
H, brt), 7.08 (1H, m), 7.18-7.27 (5H, m),
7.45 (1H, d, J = 7 Hz), 7.52 (1 H, d, J = 9.5)
Hz), 7.58 (1H, s), 7.71 (1H, m), 7.99 (1H,
d, J = 9.5 Hz)

(9) 10- [3 '-(glycylglycyl-L
-Phenylalanylglycylamino) propyloxy]
Synthesis of -7,9-trimethylene- (20S) -camptothecin hydrochloride 10- [3 '-(t-Butoxycarbonylglycylglycyl-L-phenylalanylglycylamino) propyloxy] -7,9-trimethylene- ( 273 mg of (20S) -camptothecin is dissolved in 10 ml of dioxane, and 15 ml of 18% hydrochloric acid-dioxane is added dropwise while stirring under ice cooling. The reaction mixture is stirred at room temperature, and after completion of the reaction, isopropyl ether is added, and the mixture is stirred and the precipitated powder is collected by filtration. The obtained powder was washed with ether and dried under reduced pressure. The obtained yellow powder was dissolved in water and freeze-dried to give the title compound 210 as a yellow powder.
get mg.

Melting point: 174 ° C. or higher (decomposition) Yield: 83% IR (Nujol): ν _max ^{cm −1} = 3190, 1745, 165
0 Mass: m / z = 780 ([M-Cl] ⁺ ) NMR (300 MHz, d ₆ -DMSO): δ ^TMS = 0.89
(3H, t, J = 7Hz), 1.26-1.32 (2H, m), 1.8
6-2.04 (6H, m), 2.79 (1H, dd, J = 14Hz and 10Hz), 2.98-3.05 (5H, m), 3.28-3.
36 (2H, m), 3.54-3.88 (6H, m), 4.20 (2
H, t, J = 6 Hz), 4.45-4.54 (1 H, m), 5.19
(2H, s), 5.43 (2H, s), 7.11-7.27 (5H, m),
7.35 (1H, s), 7.71 (1H, t, J = 9.5Hz), 7.9
7 (1H, t, J = 5.5 Hz), 8.03 (1H, d, J = 9.5 H)
z), 8.19 (3H, br), 8.35 (1H, t, J = 6 Hz), 8.
43 (1H, d, J = 8Hz), 8.65 (1H, t, J = 5.5H
z)

The camptothecin compound having an amino group obtained by the above synthesis was converted to carboxymethyl dextran.
(Hereinafter, CM-dextran) to condense with a water-soluble polymer having a carboxyl group to lead to a camptothecin derivative. The condensation reaction is carried out in the presence of a condensing agent such as water-soluble carbodiimide (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline). Alternatively, the reaction can be performed in a mixed solvent of water and an organic solvent.

(10) Preparation of camptothecin derivative 500 mg of CM-dextran sodium salt (degree of CM = 0.5) was dissolved in 20 ml of water, and 10- [3 '-(glycylglycyl- L-Phenylalanylglycylamino) propyloxy] -7,9-trimethylene- (20S) -camptothecin hydrochloride (50 mg) is added. 1.5 g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride was added while the reaction mixture was p
H is maintained at 6.5-7.0 with 0.1N hydrochloric acid. 10 ℃
After the reaction for 2 hours with stirring as described below, the pH is adjusted to 9 with 0.1 N sodium hydroxide. The reaction mixture was subjected to ion exchange column chromatography (BioRad AGMP).
(-50, 30 ml, Na type) and fraction 3 containing the target compound.
0 ml was added with 1.2 ml of 3M saline, and 150 ml of ethanol was added.
Pour inside. The precipitate is centrifuged, added with 20 ml of water and filtered. 0.4 ml of 3M saline was added to the filtrate, and 80 ml of ethanol was added.
Add to l with stirring to form a precipitate. The precipitate is collected by centrifugation, washed with a solvent, and dried under reduced pressure to obtain 415 mg of the camptothecin derivative represented by the above formula (13). The drug (the compound obtained by 1- (7); 10- (3′-aminopropyloxy) -7,9-trimethylene- (20S) -camptothecin hydrochloride) content determined by absorption at 380 nm was 4.
4%. Gel permeation column chromatography (GP
As a result of the analysis according to C), the average molecular weight obtained was 160,
000, polydispersity Mw / Mn is 1.57 (GPC analysis conditions: G4000SWXL (manufactured by Tosoh Corporation), 0.2 M phosphate buffer (pH 7.0)).

Example 2 Synthesis of a camptothecin derivative represented by the following formula

Embedded image (1) Synthesis of 7- (4 ′-(t-butoxycarbonylglycylglycyl-L-phenylalanylglycyl) piperazino) methyl-10,11-ethylenedioxy- (20S) -camptothecin Example 1- (8 7) -piperazinomethyl-1
From 450 mg of 0,11-ethylenedioxy- (20S) -camptothecin hydrochloride and 2 equivalents of t-butoxycarbonyl-glycylglycyl-L-phenylalanylglycine, 518 mg of the title compound is obtained as a yellow powder.

Yield: 74% IR (Nujol): ν _max ^{cm -1} = 3280,1750,165
5 Mass: m / z = 923 ([M + H] ⁺ ) NMR (300 MHz, d ₆ -DMSO): δ ^TMS = 0.89
(3H, t, J = 7.5Hz), 1.37 (9H, s), 1.85-2.
1 (2H, m), 2.3-2.6 (4H, m), 2.75 (1H, dd, J
= 14 Hz and 10 Hz), 3.05 (1 H, dd, J = 14H
z and 4.5 Hz), 3.3-3.6 (6H, m), 3.58 (1
H, dd, J = 21 Hz and 5.5 Hz), 3.74 (1H, dd,
J = 17 Hz and 5.5 Hz), 3.9-4.1 (4 H, m),
4.44 (4H, s), 4.58 (1H, m), 5.24 (2H, s),
5.42 (2H, s), 6.50 (1H, s), 6.97 (1H, t, J
= 6 Hz), 7.1-7.3 (6 H, m), 7.55 (1 H, s), 7.
77 (1H, s), 7.8-7.9 (1H, br), 8.05-8.2
(2H, m)

(2) 7- (4 '-(glycylglycyl-L-
Phenylalanylglycyl) piperazino) methyl-10,
Synthesis of 11-ethylenedioxy- (20S) -camptothecin hydrochloride In the same manner as in Example 1- (9), 7- (4 ′-(t-butoxycarbonylglycylglycyl-L-phenylalanylglycyl) piperazino ) From 478 mg of methyl-10,11-ethylenedioxy- (20S) -camptothecin, 409 mg of the title compound was obtained as a yellow powder.

Melting point: 237-239 ° C. (decomposition) IR (Nujol): ν _max ^{cm −1} = 3250,1745,165
5 Mass: m / z = 823 ([M-Cl] ⁺ ) NMR (300 MHz, d ₆ -DMSO): δ ^TMS = 0.88
(3H, t, J = 7Hz), 1.8-1.99 (2H, m), 2.79 (1
H, dd, J = 14 Hz and 10 Hz), 3.07 (1 H, dd, J
= 14 Hz and 4 Hz), 3.1-4.3 (16 H, m), 4.4
7 (4H, s), 4.55-4.70 (1H, m), 5.44 (2H,
s), 5.67 (2H, s), 7.15-7.32 (6H, m), 7.6
5 (1H, s), 8.05 (1H, s), 8.05-8.20 (3H, b
r), 8.29 (1H, br), 8.39 (1H, d, J = 8.5 Hz),
8.57 (1H, t, J = 5.5Hz)

(3) Preparation of Camptothecin Derivative In the same manner as in Example 1- (10), CM-dextran
1.2 g of sodium salt (degree of CM = 0.5) and 7- (4 '-(glycylglycyl-L-phenylalanylglycyl) piperazino) methyl-10,11-ethylenedioxy- (20
S) -camptothecin hydrochloride 168 mg
Get. Drug (7-) determined by absorption at 380 nm
Piperazinomethyl-10,11-ethylenedioxy- (2
The (OS) -camptothecin hydrochloride) content is 1.1%.
As a result of analysis by gel permeation column chromatography (GPC), the average molecular weight obtained was 169,000, and the polydispersity Mw / Mn was 1.32 (GPC analysis conditions: G40
00SWXL (manufactured by Tosoh Corporation), 0.2 M phosphate buffer (pH
7.0)).

Example 3 Synthesis of camptothecin derivative represented by the following formula

Embedded image (1) 7-N- (t-butoxycarbonylglycylglycyl-L-phenylalanylglycyl) aminomethyl-10,
Synthesis of 11-ethylenedioxy- (20S) -camptothecin 7-aminomethyl-10, as in Example 1- (8)
From 222 mg of 11-ethylenedioxy- (20S) -camptothecin hydrochloride and 2 equivalents of t-butoxycarbonylglycylglycyl-L-phenylalanylglycine, 232 mg of the title compound is obtained as a yellow powder.

Yield: 58% IR (Nujol): ν _max ^{cm -1} = 3285,1750,165
0 Mass: m / z = 854 ([M + H] ⁺ ) NMR (300 MHz, d ₆ -DMSO): δ ^TMS = 0.88
(3H, t, J = 7.5Hz), 1.35 (9H, s), 1.78-1.
94 (2H, m), 2.74 (1H, dd, J = 14 Hz and 10
Hz), 2.99 (1 H, dd, J = 14 Hz and 4.5 Hz),
3.4-3.8 (4H, m), 4.34-4.50 (1H, m), 4.
42 (4H, s), 4.66-4.82 (2H, m), 5.42 (4
H, brs), 6.50 (1H, s), 6.98 (1H, t, J = 6Hz),
7.12-7.28 (5H, m), 7.26 (1H, s), 7.56
(1H, s), 7.80 (1H, s), 7.91 (1H, br), 8.14
(1H, d, J = 7.5Hz), 8.32 (2H, t, J = 7.5H
z), 8.58 (1H, m)

(2) 7-N- (glycylglycyl-L-
(Phenylalanylglycyl) aminomethyl-10,11-
Synthesis of ethylenedioxy- (20S) -camptothecin hydrochloride In the same manner as in Example 1- (9), 7-N- (t-butoxycarbonylglycylglycyl-L-phenylalanylglycyl) aminomethyl-10, 11-ethylenedioxy-
From 203 mg of (20S) -camptothecin, 164 mg of the title compound were obtained as a yellow powder.

Melting point:> 211 ° C. (decomposition) IR (Nujol): ν _max ^{cm −1} = 3220,1745,165
5 Mass: m / z = 754 ([M-Cl] ⁺ ) NMR (300 MHz, d ₆ -DMSO): δ ^TMS = 0.88
(3H, t, J = 7Hz), 1.80-1.93 (2H, m), 2.7
7 (1 H, dd, J = 14 Hz and 10 Hz), 3.00 (1 H, d
d, J = 14 Hz and 4 Hz), 3.6-4.55 (7 H, m),
4.42 (4H, s), 4.65-4.85 (2H, m), 5.42
(2H, s), 5.45 (2H, s), 7.13-7.26 (5H, m),
7.27 (1H, s), 7.57 (1H, s), 7.83 (1H, s),
8.03-8.16 (3H, br), 8.34-8.40 (2H,
m), 8.54 (1H, br), 8.73 (2H, br)

(3) Preparation of camptothecin derivative CM-dextran sodium salt (CM conversion = 0.5)
Dissolve 772 mg in 50 ml of water and add 25 ml of DMF.
The mixture was stirred under ice-cooling to give 7- (N- (glycylglycyl-L-phenylalanylglycyl) aminomethyl) -10,11-ethylenedioxy- (20S) -camptothecin hydrochloride 10
6 mg and 2-ethoxy-1-ethoxycarbonyl-
1.57 g of 1,2-dihydroquinone are added. After overnight reaction, the reaction mixture was added to 450 ml of ethanol to form a precipitate, which was treated in the same manner as in Example 1- (10) to give 545 mg of the camptothecin derivative represented by the above formula (15) as a pale yellow powder.
Get. Drug (7-
Aminomethyl-10,11-ethylenedioxy- (20
The (S) -camptothecin hydrochloride) content is 5.5%. As a result of analysis by gel permeation column chromatography (GPC), the average molecular weight determined was 165,000, and the polydispersity Mw / Mn was 1.40 (GPC analysis conditions: G400
0SWXL (manufactured by Tosoh Corporation), 0.2 M phosphate buffer (pH
7.0)).

Example 4 Synthesis of 10- (3'-aminopropyloxy)-(20S) -camptothecin hydrochloride (1) 5- [3 '-(t-butoxycarbonylamino) propyloxy] -2-nitrobenzaldehyde Synthesis of dimethyl acetal 5-hydroxy-2-nitrobenzaldehyde 3.0 g of dimethyl acetal was dried in 50 ml of dimethylformamide
And 3.15 g of sodium iodide and 1.93 g of potassium carbonate and 3- (t-butoxycarbonylamino)
6.95 g of propyl tosylate are added. 3 at 50 ° C
After stirring for an hour, return to room temperature and add ethyl acetate. The extract is washed with saturated saline and dried over sodium sulfate.
After evaporating the solvent, the residue was separated and purified by silica gel column chromatography to obtain 5.22 g of the title compound as a pale yellow oil.
Is quantitatively obtained. IR (Neat): ν _max ^{cm -1} = 3360,1710 Mass: m / z = 393 ([M + Na] ⁺ ) NMR (300 MHz, CDCl ₃ ): δ ^TMS = 1.44 (9H,
s), 2.02 (2H, quint., J = 6 Hz), 3.33 (2H, dd,
J = 13 Hz and 6 Hz), 3.44 (6 H, s), 4.11 (2
H, t, J = 6Hz), 4.7 (1H, brs), 6.01 (1H, s),
6.90 (1 H, dd, J = 9 Hz and 3 Hz), 7.29 (1
H, d, J = 3 Hz), 7.97 (1 H, d, J = 9 Hz)

(2) Synthesis of 10- [3 ′-(t-butoxycarbonylamino) propyloxy]-(20S) -camptothecin 5- [3 ′-(t-butoxycarbonylamino) propyloxy] -2-nitro 1270 mg of benzaldehyde dimethyl acetal was dissolved in 20 ml of ethanol, and 120 mg of 10% palladium-carbon was added.
Stir for hours. The catalyst was removed by filtration from the reaction mixture, and the filtrate was added with (4S) -7,8-dihydro-4-ethyl-4-hydroxy-1H-pyrano [3,4-f] indolinidine-3,6,
300 mg of 10 (4H) -trione and 22 mg of p-toluenesulfonic acid are added and stirred overnight at room temperature. The solvent was distilled off from the reaction mixture, and the residue was separated and purified by silica gel column chromatography to obtain 204 mg of the title compound as a pale yellow powder.

Melting point: 223-224 ° C. (decomposition) Yield: 34% IR (Nujol): ν _max ^{cm −1} = 3360, 1750, 1690 Mass: m / z = 522 ([M + H] ⁺ ) NMR (300 MHz, CDCl ₃ ): δ ^TMS = 1.03 (3H,
t, J = 7.5 Hz), 1.46 (9 H, s), 1.8-2.0 (2 H,
m), 2.08 (1 H, dd, J = 12.5 Hz and 6.5 Hz),
2.10 (1H, dd, J = 12.5Hz and 6Hz), 3.40
(2H, q, J = 6.5Hz), 4.18 (2H, t, J = 6Hz),
4.82 (1H, brs), 5.24 (2H, s), 5.29 (1H, d,
J = 16 Hz), 5.73 (1 H, d, J = 16 Hz), 7.12
(1 H, d, J = 3 Hz), 7.43 (1 H, dd, J = 9 Hz and 3 Hz), 7.61 (1 H, s), 8.09 (1 H, d, J = 9 Hz),
8.20 (1H, d, J = 9Hz)

(3) 10- (3'-aminopropyloxy)
Synthesis of-(20S) -camptothecin hydrochloride 352 mg of 10- [3 '-(t-butoxycarbonylamino) propyloxy]-(20S) -camptothecin is dissolved in dry dioxane-ethanol (7 ml-1 ml) and stirred with ice cooling. Under this solution, 5 ml of 19% hydrochloric acid-dioxane is added.
After stirring the reaction mixture at room temperature, 10 ml of isopropyl ether was further added, and the precipitated powder was collected by filtration and washed to obtain 339 mg of the title compound as a yellow powder.

Melting point: 214-218 ° C. (decomposition) IR (Nujol): ν _max ^{cm −1} = 3470,3280,174
5 Mass: m / z = 422 ([M-Cl] ⁺ ) NMR (300 MHz, d ₆ -DMSO): δ ^TMS = 0.89
(3H, t, J = 7.5Hz), 1.80-1.95 (2H, m), 2.
10-2.22 (2H, m), 2.96-3.10 (2H, m), 4.
27 (2H, t, J = 6Hz), 5.25 (2H, s), 5.42 (2
H, s), 7.29 (1H, s), 7.49-7.55 (2H, m), 8.
08 (1H, d, J = 10 Hz), 8.19 (3H, brs), 8.54
(1H, s)

Example 5 Synthesis of 10- [3 '-(glycyl-glycyl-L-phenylalanyl-glycylamino) propyloxy]-(20S) -camptothecin hydrochloride (1) 10- [3'-(t- Butoxycarbonyl-glycyl-
Synthesis of glycyl-L-phenylalanylglycylamino) propyloxy]-(20S) -camptothecin
325 mg of (3'-aminopropyloxy)-(20S) -camptothecin hydrochloride was added to 10 ml of dry dimethylformamide.
And 4 equivalents of N-hydroxysuccinimide, 2 equivalents of diisopropylethylamine, 2 equivalents of Nt-butoxycarbonyl-glycyl-glycyl-L-phenylalanylglycine and 4 equivalents of An equivalent amount of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is added. After stirring the reaction mixture at room temperature overnight, the solvent is distilled off. The residue is separated and purified by silica gel column chromatography to quantitatively obtain 597 mg of the title compound as a yellow powder.

IR (Nujol): ν _max ^{cm -1} = 3280,17
50,1660 Mass: m / z = 840 ([M + H] ⁺ ) NMR (300 MHz, d ₆ -DMSO): δ ^TMS = 0.88
(3H, t, J = 7Hz), 1.36 (9H, s), 1.8-2.04
(4H, m), 2.72-2.84 (1H, m), 3.00-3.12
(1H, m), 3.24-3.36 (2H, m), 3.50-3.80
(6H, m), 4.18 (2H, t, J = 6Hz), 4.44-4.5
4 (1H, m), 5.24 (2H, s), 5.42 (2H, s), 6.50
(1H, s), 6.99 (1H, t, J = 6Hz), 7.12-7.2
7 (5H, m), 7.28 (1H, s), 7.48-7.55 (1H,
m), 7.50 (1H, s), 7.88-7.96 (1H, m), 8.0
7 (1H, d, J = 9Hz), 8.12-8.36 (2H, m), 8.
51 (1H, s)

(2) 10- [3 '-(glycyl-glycyl-
L-phenylalanylglycylamino) propyloxy]
Synthesis of-(20S) -camptothecin hydrochloride Example 4- (10- (3 '-(t-Butoxycarbonyl-glycyl-glycyl-L-phenylalanylglycylamino) propyloxy)-(20S) -camptothecin from Example 580 mg Work-up as in 3) to give 438 mg of the title compound as a yellow powder.

Yield: 82% Melting point: 194-199 ° C. (decomposition) IR (Nujol): ν _max ^{cm −1} = 3190, 1745, 165
0 Mass: m / z = 740 ([M-Cl] ⁺ ) NMR (300 MHz, d ₆ -DMSO): δ ^TMS = 0.88
(3H, t, J = 7Hz), 1.80-2.03 (4H, m), 2.7
9 (1 H, dd, J = 14 Hz and 10 Hz), 3.05 (1 H, d
d, J = 14 Hz and 5 Hz), 3.2-3.5 (2 H, m), 3.
52-3.62 (2H, m), 3.62-3.83 (4H, m), 4.
19 (2H, t, J = 6Hz), 4.48-4.58 (1H, m),
5.25 (2H, s), 5.42 (2H, s), 6.5 (1H, brs),
7.13-7.26 (5H, m), 7.28 (1H, s), 7.49-
7.55 (1H, m), 7.50 (1H, s, J = 9.5Hz), 7.9
3 (1H, t, J = 6Hz), 8.0-8.14 (4H, m), 8.3
2-8.41 (2H, m), 8.51 (1H, s), 8.56 (1H,
t, J = 5.5Hz)

Example 6 Synthesis of camptothecin derivative represented by the following formula

Embedded image CM-dextran sodium salt (CM degree = 0.5)
513 mg was dissolved in 50 ml of water, and 10- [3 ′-(glycyl-glycyl-L-phenylalanyl-glycylamino) propyloxy]-(20
77 mg of S) -camptothecin hydrochloride and 1.54 g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride are added. The mixture is stirred for 2 hours while maintaining the internal temperature at 10 ° C. or lower and the pH at 6.0 to 6.5 using 0.2N hydrochloric acid. Ion exchange column chromatography of the reaction mixture
(BioRad AGMP-50, 30 ml, Na type), the fraction containing the target substance was filtered, ethanol was added to the filtrate, the precipitate was collected, washed, and dried under reduced pressure to obtain a pale yellow powdery complex. 492 mg are obtained. The drug (10- (3′-aminopropyloxy)-(20
S) -Camptothecin hydrochloride) content is 2.8%. As a result of analysis by gel filtration column chromatography (GPC), the average molecular weight obtained was 179,000 and the polydispersity Mw / Mn was 1.42 (GPC analysis conditions: G400
0SWXL (manufactured by Tosoh Corporation), 0.2 M phosphate buffer (pH =
7.0)).

Example 7 Synthesis of camptothecin derivative represented by the following formula

Embedded image (1) (9S) -1- (t-butoxycarbonylglycylglycyl-L-phenylalanylglycylamino) -9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H , 12H-benzo [de] pyrano [3 ',
4 ′: 6,7] Indolizino [1,2-b] quinoline-10,1
Synthesis of 3 [9H, 15H] -dione (9S) -1-amino-9 in the same manner as in Example 1- (8).
-Ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H, 12H-benzo [de] pyrano
[3 ', 4': 6,7] Indolizino [1,2-b] quinoline-1
166 mg of 0.13 (9H, 15H) -dione hydrochloride and 247 mg of the title compound as a pale yellow amorphous solid were obtained from 2 equivalents of t-butoxycarbonylglycylglycyl-L-phenylalanylglycine.

Yield: 82% IR (Nujol): ν _max ^{cm -1} = 3290,1710,165
5 Mass: m / z = 854 ([M + H] ⁺ ) NMR (300 MHz, d ₆ -DMSO): δ ^TMS = 0.87
(3H, t, J = 7Hz), 1.37 (9H, s), 1.8-1.95
(2H, m), 2.05-2.3 (1H, m), 2.42 (3H, s),
2.5-2.85 (2H, m), 2.9-3.1 (1H, m), 3.1
5-3.4 (2H, m), 3.5-3.8 (6H, m), 4.4-4.
55 (1H, m), 5.26 (2H, s), 5.42 (2H, s), 5.5
5-5.65 (1H, m), 6.53 (1H, s), 6.99 (1H,
t, J = 5Hz), 7.1-7.3 (5H, m), 7.32 (1H, s), 7.
81 (1H, d, J = 11Hz), 7.8-7.95 (1H, m),
8.1-8.2 (1H, m), 8.3-8.4 (1H, m), 8.4
8.5 (1H, m)

(2) (9S) -1- (glycylglycyl-L
-Phenylalanylglucylamino) -9-ethyl-5
-Fluoro-2,3-dihydro-9-hydroxy-4-
Methyl-1H, 12H-benzo [de] pyrano [3 ', 4': 6,
7] Indolizino [1,2-b] quinoline-10,13 [9H,
Synthesis of 15H] -dione Compound 2 obtained in (1) above in the same manner as in Example 1- (9)
From 20 mg, 193 mg of the title compound were obtained as a pale yellow powder. Melting point:> 165 ° C (decomposition) IR (Nujol): ν _max ^{cm -1} = 3350,1745,166
0.11615 Mass: m / z = 754 ([M-Cl + H] ⁺ ) NMR (300 MHz, d ₆ -DMSO): δ ^TMS = 0.87
(3H, t, J = 7Hz), 1.80-1.94 (2H, m), 2.0
8-2-2.7 (2H, m), 2.41 (3H, s), 2.77 (1H, d
d, J = 13 Hz, 9 Hz), 3.01 (1 H, dd, J = 13 Hz,
5Hz), 3.15-3.28 (2H, m), 3.5-3.91 (6
H, m), 4.45-4.56 (1H, m), 5.25 (2H, s), 5.
41 (1H, d, J = 13Hz), 5.42 (1H, d, J = 13H)
z), 5.57 (1H, m), 7.12-7.30 (5H, m), 7.3
2 (1H, s), 7.80 (1H, d, J = 11 Hz), 8.0-8.
2 (3H, br), 8.32 (1H, d, J = 7Hz), 8.43 (1
H, t, J = 5.5 Hz), 8.50-8.62 (2 H, m)

(3) Preparation of camptothecin derivative CM-dextran sodium salt (CM conversion = 0.6)
5) The same treatment as in Example 3- (3) was carried out using 2000 mg and the compound (170 mg) obtained in the above (2) to obtain 1803 mg of the desired camptothecin derivative as a pale yellow powder. 37
Drug ((9S) -1-amino-9-ethyl-5-fluoro-2,3-dihydro-9) determined by absorption at 6 nm
-Hydroxy-4-methyl-1H, 12H-benzo [de]
The content of pyrano [3 ', 4': 6,7] indolizino [1,2-b] quinoline-10,13 [9H, 15H] -dione hydrochloride) is 3.
0%. Gel filtration column chromatography (GP
As a result of the analysis according to C), the average molecular weight obtained was 187,
000, and the polydispersity Mw / Mn is 1.54 (GPC analysis conditions: G4000SWXL (manufactured by Tosoh Corporation), 0.2 M phosphate buffer (pH = 7.0)).

Examples 8 to 24 In the same manner as in Example 1-6, the compounds shown in Table 1 below are obtained.

[Table 1] [CM-Pullulan ・ Na is carboxymethyl pullulan ・
Represents sodium salt]

Examples 25 to 41 The compounds shown in Table 2 below were obtained in the same manner as in Example 1-6.

[Table 2]

Examples 42 to 59 The compounds shown in Table 3 below were obtained in the same manner as in Example 1-6.

[Table 3]

Examples 60 to 76 The compounds shown in Table 4 below were obtained in the same manner as in Examples 1-6.

[Table 4]

Examples 77 to 93 The compounds shown in Table 5 below were obtained in the same manner as in Examples 1-6.

[Table 5]

Examples 94 to 110 The compounds shown in Table 6 below were obtained in the same manner as in Examples 1-6.

[Table 6]

Reference Example 1 (1) Dextran [Dextran T- manufactured by Pharmacia]
110, average molecular weight: 100,000 (GPC) method] 29 g
Is dissolved in 290 ml of water. To this solution is added 1.45 g of sodium borohydride at 0-5 ° C and stirred at 5 ° C overnight. Acetic acid was added dropwise to the reaction mixture to adjust the pH to 5, and the mixture was further stirred at room temperature for 3 hours. The pH is adjusted to 7 with 2N sodium hydroxide and, with vigorous stirring, 1.2 l of ethanol are added. After allowing to stand to precipitate insolubles, the supernatant is removed by decantation, and the residue is centrifuged.
The residue is dissolved in 0.5 liter of water and lyophilized to give 26.3 g of a white powder. (2) Dissolve 50 g of the white powder obtained in the above (1) in 500 ml of water, and add 200 g of sodium hydroxide to this solution under ice-cooling.
And stir for 30 minutes. After returning the reaction mixture to room temperature, a solution of monochloroacetic acid (110 g) in water (150 ml) was added dropwise, and the mixture was stirred at 40 ° C. for 18 hours. Reaction mixture at 10 ° C
Cool below and adjust to pH 8-9 with acetic acid. While stirring the reaction mixture vigorously, 8 liters of methanol are added to precipitate insolubles. The insolubles are collected by filtration, dissolved in 5 liters of pure water, and desalted by ultrafiltration. The resulting solution was concentrated under reduced pressure, the concentrated solution was filtered, ethanol was added, the precipitate was collected by filtration, washed with aqueous ethanol and acetone, and dried under reduced pressure at room temperature and 50 ° C. to obtain carboxymethyl dextran ( CM-dextran) sodium salt
[Degree of carboxymethylation (neutralization titration method): 0.5] 50.2 g
Get.

Reference Example 2 The same procedure as in Reference Example 1 was carried out except that the amount of monochloroacetic acid used was changed to obtain a C-carboxymethylated compound having a degree of carboxymethylation of 0.65.
M-dextran sodium salt is obtained.

Continued on the front page (51) Int.Cl. ⁶ Identification code FI C07D 491/22 C07D 491/22 C08B 37/02 C08B 37/02

Claims (14)

[Claims] 1. A compound of the general formula [Wherein, R ^{1 to} R ⁵ are (A) two adjacent R ^{1 to} R ⁵ are bonded to each other to form an alkylene group, or both are hydrogen atoms, and the remaining R ¹ out of ^{1 to} R ⁵
One is a -X _n -Alk _m -R ^6, the other two are hydrogen atom,
An alkyl group or a halogen atom, or (B)
Among R ¹ to R ^5, two adjacent but an alkylene group formed by combining with each other, -X _n -Alk _m -R ⁶ are substituted to any carbon atom of the alkylene group, R ¹ ~ the remaining three are hydrogen atoms R ^5, represents an alkyl group or a halogen atom, (a) and one or two methylene groups in the alkylene group in (B) is -O -, - S- or - X may be replaced with NH-, X is -O- or -NH-, Alk is an alkylene group, R ⁶
Is —NH ₂ , Or m and n are both 0 or 1, or m is 1 and n is 0], and a polysaccharide having a carboxyl group is bonded via an amino acid or a peptide. Or a pharmacologically acceptable salt thereof. 2. A part or all of a carboxyl group of a polysaccharide and an amino group of an amino acid or a peptide form an acid amide bond, and all or a part of the carboxyl group of the amino acid or the peptide and R ^{6 of the} compound [I] The compound according to claim 1, which is formed by an acid amide bond or an ester bond, or a pharmaceutically acceptable salt thereof. 3. The carboxyl group of the polysaccharide is partially or entirely bonded to the N-terminal amino group of the amino acid or peptide by an acid amide bond, and the C-terminal carboxyl group of this amino acid or peptide is linked to R ^{6 of} compound [I]. 3. The compound according to claim 2, which is formed by an acid amide bond or an ester bond, or a pharmaceutically acceptable salt thereof. 4. The compound of formula [I] wherein R ⁶ has the formula: —NH ₂ or The compound according to claim 3, wherein the polysaccharide having a carboxyl group is carboxymethylated dextran or pullulan, and both components are bonded via a peptide, or a pharmaceutically acceptable salt thereof. 5. (1) R ¹ and R ² are bonded to form a trimethylene group, R ³ is a 3-aminopropyloxy group, R ⁴ and R ⁵ are hydrogen atoms, (2) R ¹ is a piperazinomethyl group, R ² And R ⁵
Is a hydrogen atom, R ³ and R ⁴ are bonded to an ethylenedioxy group, (3) R ¹ is an aminomethyl group, R ² and R ⁵ are a hydrogen atom, and R ³ and R ⁴ are bonded to an ethylenedioxy group. , (4) R
¹ , R ² , R ⁴ and R ⁵ are a hydrogen atom, R ³ is a 3-aminopropyloxy group, (5) an amino-substituted trimethylene group in which R ¹ and R ² are bonded, R ³ is a methyl group, and R ⁴ is Fluorine atom, R ⁵
Is a hydrogen atom, or (6) R ¹ , R ³ , R ⁴ and R ⁵ are a hydrogen atom, and R ² is an amino group, or a pharmaceutically acceptable salt thereof. 6. The method according to claim 6, wherein the peptide is glycyl-glycyl-L or D-phenylalanyl-glycine, glycyl-glycyl, glycyl-glycyl-glycine, glycyl-glycyl-glycyl-glycine, glycyl-glycyl-glycyl-glycyl-glycine, The compound according to claim 5, which is L or D-phenylalanyl-glycine or L or D-leucyl-glycine, or a pharmaceutically acceptable salt thereof. 7. The method according to claim 7, wherein the peptide is glycyl-glycyl-L-
The compound according to claim 6, which is phenylalanyl-glycine, or a pharmaceutically acceptable salt thereof. 8. The compound according to claim 6, wherein said peptide is glycyl-glycine, or a pharmaceutically acceptable salt thereof. 9. The compound according to claim 6, wherein the peptide is glycyl-glycyl-glycine, or a pharmaceutically acceptable salt thereof. 10. The compound according to claim 6, wherein the peptide is glycyl-glycyl-glycyl-glycine, or a pharmaceutically acceptable salt thereof. 11. The compound according to claim 6, wherein the peptide is L or D-phenylalanyl-glycine, or a pharmaceutically acceptable salt thereof. 12. The polysaccharide having a degree of carboxymethylation of 0.5.
12. The compound according to claim 7, 8, 9, 10 or 11, or a pharmaceutically acceptable salt thereof, which is 3 or more and 0.8 or less. 13. A compound of the general formula [Wherein, R ^{1 to} R ⁵ are (A) two adjacent R ^{1 to} R ⁵ are bonded to each other to form an alkylene group, or both are hydrogen atoms, and the remaining R ¹ out of ^{1 to} R ⁵
One is a -X _n -Alk _m -R ^6, the other two are hydrogen atom,
An alkyl group or a halogen atom, or (B)
Among R ¹ to R ^5, two adjacent but an alkylene group formed by combining with each other, -X _n -Alk _m -R ⁶ are substituted to any carbon atom of the alkylene group, R ¹ ~ the remaining three are hydrogen atoms R ^5, represents an alkyl group or a halogen atom, (a) and one or two methylene groups in the alkylene group in (B) is -O -, - S- or - X may be replaced with NH-, X is -O- or -NH-, Alk is an alkylene group, R ⁶
Is —NH ₂ , Or m and n are both 0 or 1 or m is 1 and n is 0], and a camptothecin compound obtained by bonding an amino acid or a peptide to an amino group or a carboxyl group. When the group is protected, the protecting group is removed, and then reacted with a polysaccharide having a carboxyl group, and if necessary, the product is converted into a pharmaceutically acceptable salt. And a polysaccharide having a carboxyl group bonded via an amino acid or a peptide, or a pharmaceutically acceptable salt thereof. 14. A compound of the general formula [Wherein, R ^{1 to} R ⁵ are (A) two adjacent R ^{1 to} R ⁵ are bonded to each other to form an alkylene group, or both are hydrogen atoms, and the remaining R ¹ out of ^{1 to} R ⁵
One is a -X _n -Alk _m -R ^6, the other two are hydrogen atom,
An alkyl group or a halogen atom, or (B)
Among R ¹ to R ^5, two adjacent but an alkylene group formed by combining with each other, -X _n -Alk _m -R ⁶ are substituted to any carbon atom of the alkylene group, R ¹ ~ the remaining three are hydrogen atoms R ^5, represents an alkyl group or a halogen atom, (a) and one or two methylene groups in the alkylene group in (B) is -O -, - S- or - X may be replaced with NH-, X is -O- or -NH-, Alk is an alkylene group, R ⁶
Is —NH ₂ , Or m and n are both 0 or 1, or m is 1 and n is 0] with an amino acid or a peptide, and the amino group or When the carboxyl group is protected, the protecting group is removed and, if desired, the product is converted into a salt. The compound [I] and an amino acid or a peptide are combined with a camptothecin compound or a salt thereof. Recipe.