HK1105419B - Camptothecin derivatives and their uses - Google Patents

Abstract

The present invention provides a class of new camptothecin derivatives having the structure of the following formula and having anti-tumor activity and water solubility, their uses, and the pharmaceutical composition containing the same. The compounds of the present invention have good anti-tumor activity and water solubility, and they have good prospect of being developed as medicines.

Description

Camptothecin derivatives and application thereof

Technical Field

The invention relates to the field of medicinal chemistry, in particular to a novel water-soluble camptothecin derivative with antitumor activity, application thereof and a pharmaceutical composition containing the compound.

Background

Camptothecin (CPT, J.Am.chem.Soc., 1966, 88, 3888) is extracted and separated from Camptotheca acuminata by Wall, etc., and is pyrrole [3, 4-b ]]Quinoline alkaloid with the structural formulaThe structure of the compound contains five rings, 20 positions of the E rings have an S-type chiral center, and a lactone structure is arranged beside the S-type chiral center. Camptothecin has certain curative effect on gastric cancer, rectal cancer and the like, but has poor water solubility and great toxic and side effects, so clinical research is stopped.

In order to obtain the CPT derivative with high activity and low toxicity, a large amount of structural transformation is carried out, a plurality of CPT compounds are synthesized, and a good effect is achieved. Research has found that the introduction of an appropriate group at position 9 of 10-hydroxycamptothecin can increase antitumor activity, such as topotecan which is currently on the market.

Although CPT compounds are alkaloids, the CPT compounds do not have water solubility after being salified by common inorganic bases and organic bases. In order to solve the problem of water solubility, two general schemes are provided, namely, groups which can form salts and have water solubility are introduced into CPT compounds, such as some amino groups, and the case is that topotecan; another method is to introduce temporary groups containing water solubility into CPT compounds, and the water-soluble groups of the compounds are dissociated in vivo. The second case is Irinotecan (compound 4, Irinotecan, CPT-11), a water-soluble CPT-like drug.

The inventor of the invention surprisingly found that a compound obtained by introducing a simple alkyl group into the 9-position of 10-hydroxycamptothecin (shown as the following structural formula II, wherein R1 is H, C) in the process of screening antitumor drugs₁～C₄Alkyl, C with branched chain₁～C₄Alkyl, or C substituted by hydroxy and/or amino₁～C₄Alkyl) has good antitumor activity, some compounds have good therapeutic effect on solid tumor of tumor-bearing nude mouse, and haveThe higher therapeutic index shows the prospect of the compound in the development of drugs.

However, the compound having the structure shown in formula II also has the problem of poor water solubility as described above.

Disclosure of Invention

Therefore, the object of the present invention is to provide a new class of camptothecin derivatives having high antitumor activity and being soluble in water based on the above compounds of formula II.

Another object of the present invention is to provide a pharmaceutical composition comprising the above camptothecin derivative.

The invention also aims to provide the application of the camptothecin derivative in preparing a medicament for treating tumors.

The invention also aims to provide application of the pharmaceutical composition in preparing a medicament for treating tumors.

The invention provides a compound with a structure shown in a structural formula I, an isomer, a counterpart or a medicinal salt thereof,

wherein R is₁Is H, C₁～C₄Alkyl, vinyl, hydroxy, amino, C with a branched chain₁～C₄Alkyl, or C substituted by hydroxy and/or amino₁～C₄An alkyl group. Specifically, R₁May be a hydrogen atom; methyl, ethyl, propyl, allylAlkyl, butyl, and their possible isomers, such as isopropyl, isobutyl; and hydroxymethyl, hydroxyethyl, hydroxypropyl, aminomethyl, aminoethyl, aminopropyl and the like.

Examples of the pharmaceutically acceptable salts are salts of an amine group having basicity in the piperidine group introduced at the 10-position hydroxyl group with a pharmaceutically acceptable inorganic acid or organic acid, which can make the drug water-soluble. For example, hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, trifluoroacetic acid, trifluoromethanesulfonic acid.

The compounds of the invention are preferably:

10- ((4' -piperidinylpiperidine) carbonyloxy) -9-methylcamptothecin;

10- ((4' -piperidinylpiperidine) carbonyloxy) -9-ethylcamptothecin;

10- ((4' -piperidinylpiperidine) carbonyloxy) -9-propylcamptothecin;

10- ((4' -piperidinylpiperidine) carbonyloxy) -9-allylcamptothecin;

10- ((4' -piperidinylpiperidine) carbonyloxy) -9-isopropylcamptothecin;

10- ((4' -piperidinylpiperidine) carbonyloxy) -9-n-butylcamptothecin;

10- ((4' -piperidinylpiperidine) carbonyloxy) -9-isobutylcamptothecin;

10- ((4' -piperidinylpiperidine) carbonyloxy) -9-hydroxymethylcamptothecin;

10- ((4' -piperidinylpiperidine) carbonyloxy) -9-hydroxyethyl camptothecin; or

10- ((4' -piperidinopiperidine) carbonyloxy) -9-aminomethylcamptothecin.

The compounds of the invention can be synthesized in the same manner as irinotecan is synthesized as reported in the prior art, and these methods are reported in many references (e.g., chem. pharm. Bull, 1991, 39, 2574).

The specific synthetic method can be carried out according to the following two routes, in scheme 1, the structural formula is shown inPyrrole [3, 4-b ] of]Quinoline alkaloid reacts with phosgene or solid phosgene to obtain acyl chloride compound 6, and then reacts with piperidyl piperidine (compound 7) to obtain the product.

In scheme 2, the piperidyl piperidine is first reacted with phosgene or phosgene solids to give the chloroformate amide 8, which is then reacted with the pyrrolo [3, 4-b ] quinoline alkaloid described in scheme 1 to give the product.

Both routes employ conventional organic synthesis reactions, and can be performed by the average professional with reference to the relevant synthetic literature, and need not be described in detail herein.

The pharmaceutically acceptable salts of the compounds of the present invention can be obtained by conventional methods and will not be described in detail herein. The chemical properties of the compounds of the invention allow them to be formulated as water-soluble salts.

The invention also provides a pharmaceutical composition comprising a therapeutically effective dose of the above compound and conventional pharmaceutical excipients. An "effective amount" is an amount of the compound sufficient to significantly ameliorate the condition without causing serious side effects. The safe and effective dose of the compound will be determined by the age, body weight, therapeutic indication, route of administration, course of treatment and any relevant therapeutic considerations of the subject being treated. The pharmaceutic adjuvant comprises a pharmaceutically acceptable carrier, namely' pharmacyThe above acceptable carrier "means: one or more compatible solid or liquid fillers or excipients which are suitable for human use and must be of sufficient purity and of sufficiently low toxicity. By "compatible" is meant herein that the components of the composition are capable of intermixing with and with the compounds of the present invention without significantly diminishing the efficacy of the compounds. Examples of pharmaceutically acceptable carrier moieties are sugars (e.g. glucose, sucrose, lactose, etc.), starches (e.g. corn starch, potato starch, etc.), celluloses and derivatives thereof (e.g. sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, microcrystalline cellulose, etc.), acrylics, sodium polyacrylate, povidone, polyethylene glycol, polyoxyethylene monostearate, gelatin, colloidal silica, talc, stearic acid, magnesium stearate, calcium sulfate, vegetable oils (e.g. soybean oil, sesame oil, peanut oil, olive oil, etc.). It can also be emulsifier (such as Tween)) Wetting agents (e.g., sodium lauryl sulfate), plasticizers (e.g., dibutyl sebacate), coloring agents, flavoring agents, stabilizers, preservatives, pyrogen-free water, and the like. The choice of carrier for the compositions of the invention will depend on the mode of administration of the compound and, in addition, one of ordinary skill in the art can select a carrier suitable for a particular mode of administration in light of the prior art.

The invention also provides a preparation form of the pharmaceutical composition, and the preparation form can be suitable for oral administration, intravenous injection, intramuscular injection and the like, such as powder injection, tablets, capsules and the like.

The compound of the invention has anti-tumor activity, therefore, the compound of the invention and the pharmaceutical composition containing the compound can be used for preparing drugs for treating tumors, further used for treating tumors and can also be used for interventional therapy of tumors.

The compound of the invention has good anti-tumor activity and water solubility, and the compound has excellent prospect in drug development.

Drawings

FIG. 1 shows that CPT-4 active metabolites inhibit Topo I unwinding pBR 322.

FIG. 2 shows the effect of CPT-4 active metabolites on multidrug resistance.

FIG. 3 shows the experimental therapeutic effect of CPT-4 on human colon cancer nude mouse graft tumor HCT-116.

FIG. 4 shows the experimental therapeutic effect of CPT-4 on human lung cancer nude mouse transplantable tumor SPC-A4.

Detailed Description

Preparation examples

Preparation example 110 preparation of- ((4' -Piperidinylpiperidine) carbonyloxy) -9-allylcamptothecin (CPT-4)

1g (1.25 equiv.) of piperidinylpiperidine chloroformate (Compound 8) is dissolved in 70mL of methylene chloride, 10-hydroxy-9-allylcamptothecin (1g, 1 equiv.) is dissolved in 70mL of anhydrous pyridine, the methylene chloride solution is added under cooling, after the reaction is finished, the solvent is evaporated under reduced pressure, and the residue is chromatographed on a silica gel column to obtain 1.25g of CPT-4 yellow solid.¹HNMR(DMSO-d₆) (ppm)：1.01(3H，t)，1.58～1.90(10H，m)，1.80～1.99(2H，m)，2.89(4H，b)，3.09(1H，b)，3.71(2H，d)，4.45(2H，dd)，4.94(1H，dd)，5.11(1H，dd)，5.14(2H，s)，5.15(1H，d)，5.66(1H，d)，6.00(1H，m)，7.47(1H，d)，7.65(1H，s)，8.11(1H，d)，8.51(1H，s)。

Preparation example 210 preparation of- ((4' -Piperidinylpiperidine) carbonyloxy) -9-ethylcamptothecin (CPT-2)

Piperidinylpiperidine chloroformate amide (Compound 8)0.95g (1.25 equiv.) was dissolved in 70mL of methylene chloride, and 10-hydroxy-9-ethylcamptothecin (1g, 1 equiv.) was dissolved in 70mL of anhydrous pyridineAnd adding the dichloromethane solution into pyridine under the cooling condition, evaporating the solvent under reduced pressure after the reaction is finished, and performing silica gel column chromatography on the residue to obtain 1.24g of CPT-2 yellow solid.¹HNMR(DMSO-d₆)(ppm)：1.01(3H，t)，1.20(3H，t)，1.58～1.90(2H，m)，1.80～1.99(2H，m)，2.89(4H，b)，3.09(1H，b)，3.21(2H，q)，4.45(2H，dd)，5.14(2H，s)，5.15(1H，d)，5.66(1H，d)，6.00(1H，m)，7.47(1H，d)，7.65(1H，s)，8.11(1H，d)，8.67(1H，s)。

Preparation example 310 preparation of- ((4' -Piperidinylpiperidine) carbonyloxy) -9-propylcamptothecin (CPT-3)

1g (1.25 equiv.) of piperidinylpiperidine chloroformate (Compound 8) is dissolved in 70mL of methylene chloride, 10-hydroxy-9-propylcamptothecin (1g, 1 equiv.) is dissolved in 70mL of anhydrous pyridine, the methylene chloride solution is added under cooling, after the reaction is finished, the solvent is evaporated under reduced pressure, and the residue is chromatographed on a silica gel column to obtain 1.17g of CPT-3 as a yellow solid.¹HNMR(DMSO-d₆)(ppm)：1.01(3H，t)，1.12(3H，t)，1.59(2H，m)，1.82～1.90(2H，m)，1.80～1.99(2H，m)，2.89(4H，b)，3.09，3.22(2H，t)，(1H，b)，4.45(2H，dd)， 5.14(2H，s)，5.15(1H，d)，5.66(1H，d)，6.00(1H，m)，7.47(1H，d)，7.65(1H，s)，8.11(1H，d)，8.51(1H，s)。

Test examples

The following pharmacological test was conducted using the compound of the present invention CPT-4 prepared as described above.

Test example 1 inhibition of TOPO I by cell-free System

The TOPO I mediated negative supercoiled PBR322 unwinding experiment is used to detect the effect of CPT-4 active metabolite on TOPO I enzyme activity. In cell-free system experiments (FIG. 1, wherein RLX: unwinding DNA; SC: supercoiled DNA), the CPT-4 active metabolite was able to inhibit the unwinding of supercoiled DNA by Topo I, and at the same concentration, the inhibition of CPT-4 active metabolite was stronger than that of CPT, TPT (topotecan) and SN38 (technical drug of irinotecan).

Test example 2 in vitro antitumor Activity

1. The sulforhodamine B protein staining method (SRB method) is used for detecting the effect of the compound on inhibiting the proliferation of tumor cells. The results showed that CPT-4 active metabolite was effective in inhibiting tumor cell proliferation at lower concentrations (Table 1), which was IC-resistant to thirteen tumor cells₅₀Average values of (115.2nM) were lower than the control compounds TPT (378.6nM), SN38(218.5nM) and 9-nitrocamptothecin (9-NC) (167.0 nM). Meanwhile, the compound has selectivity on tumor cell strains with different tissue sources. Among them, lung cancer, colon cancer and breast cancer cell lines are sensitive to CPT-4 active metabolites, while their effects on liver cancer, stomach cancer and ovarian cancer are relatively weak (see Table 1 for results).

TABLE 1 inhibition of tumor cell proliferation by CPT-4 active metabolites

2. The multidrug resistance cell strain K562/A02 and the parent K562 cell are selected to evaluate the multidrug resistance effect of the CPT-4 active metabolite. IC of Doxorubicin on K562 and K562/A02 cells₅₀0.493 and 69.141 μ M, respectively, and a Resistance Factor (RF) of 140.24. The result shows that the CPT-4 active metabolite has equivalent cytotoxicity to two cells, presents obvious multi-drug resistance effect and is stronger than TPT and SN38 (the result is shown in figure 2, wherein, (A) IC50 to MDR K562/A02 and parent K562 cells thereof, (B) drug resistance factor).

Experimental example 3 in vivo antitumor Activity study

HCT for human colon cancer116 transplantable tumor or human lung cancer SPC-A4 transplantable tumor is inoculated to the armpit of a nude mouse until the tumor volume reaches 100-200 mm³At the time, the drugs were administered in random cages, and CPT-4 or physiological saline was administered intravenously at different concentrations three times per week. The results show that the compound can obviously inhibit the growth of transplanted tumors, and the action intensity is equivalent to that of CPT-11 (water-soluble CPT drug irinotecan), and the results are shown in tables 2 and 3.

TABLE 2 Experimental therapeutic Effect of CPT-4 on human Colon cancer nude mouse graft tumor HCT-116

TABLE 3 Experimental therapeutic Effect of CPT-4 on human Lung cancer nude mouse transplantable tumor SPC-A4

Note: one of the CPT-415 mg/kg groups had complete tumor regression in nude mice.

Claims (6)

1. A compound with a structure shown in a structural formula I or a medicinal salt thereof,

wherein R is₁Is C₁～C₄Alkyl, vinyl or C substituted by hydroxy and/or amino₁～C₄An alkyl group.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

10- ((4' -piperidinylpiperidine) carbonyloxy) -9-ethylcamptothecin;

10- ((4' -piperidinylpiperidine) carbonyloxy) -9-propylcamptothecin;

10- ((4' -piperidinopiperidine) carbonyloxy) -9-allylcamptothecin.

3. An antitumor pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of claims 1-2 or a pharmaceutically acceptable salt thereof and conventional pharmaceutical excipients.

4. The antitumor pharmaceutical composition of claim 3, wherein said pharmaceutical composition is formulated as an oral or injectable dosage form.

5. Use of a compound according to any one of claims 1-2, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a neoplastic disease.

6. Use of a pharmaceutical composition according to claim 3 for the preparation of a medicament for the treatment of a tumor disease.