CN111407760B

CN111407760B - Pharmaceutical composition for treating infantile osteosarcoma

Info

Publication number: CN111407760B
Application number: CN202010302183.8A
Authority: CN
Inventors: 袁悦; 陆明; 查巍; 褚祥军; 刘永; 袁毅; 孙军
Original assignee: Anhui provincial children's hospital
Current assignee: Anhui provincial children's hospital
Priority date: 2020-04-16
Filing date: 2020-04-16
Publication date: 2021-05-04
Anticipated expiration: 2040-04-16
Also published as: CN111407760A

Abstract

The present invention relates to a pharmaceutical composition for the treatment of pediatric osteosarcoma comprising a) 2-methyl-2- (4- {5- [3- (methylsulfonyl) phenyl ] -1H-pyrrolo [2,3-b ] pyridin-3-yl } phenyl) propanoic acid or a pharmaceutically acceptable salt thereof and b) a pharmaceutically acceptable excipient. The invention also relates to the use of 2-methyl-2- (4- {5- [3- (methylsulfonyl) phenyl ] -1H-pyrrolo [2,3-b ] pyridin-3-yl } phenyl) propanoic acid or a pharmaceutically acceptable salt thereof in the manufacture of a pharmaceutical composition for the treatment of pediatric osteosarcoma.

Description

Pharmaceutical composition for treating infantile osteosarcoma

Technical Field

The invention relates to the technical field of medicines, and particularly relates to a pharmaceutical composition for treating infantile osteosarcoma.

Background

Sarcoma (sarcoma) refers to a malignant tumor that originates in mesenchymal tissue. Osteosarcoma is a common sarcoma, is usually developed from mesenchymal cell lines in children and young people, and is most commonly found at the two ends of long bones of limbs, especially at the lower end of femur, upper end of tibia and upper end of humerus. The osteosarcoma has short course and rapid growth, and can generate hematogenous metastasis in early stage. Osteosarcoma grows in cortex initially, can gradually develop into a bone marrow cavity, sometimes breaks through periosteum and infiltrates into surrounding soft tissues, and pathological fracture is easily caused. Generally speaking, osteosarcoma has high malignancy degree, and is often accompanied with adverse consequences such as lung metastasis, pain-avoiding claudication, muscle atrophy, joint movement limitation, amputation, death and the like, and the metastasis rate, the recurrence rate and the death rate are high.

Currently, the treatment of osteosarcoma is still based on surgical treatment, with appropriate combination of radiotherapy and chemotherapy. Radical operation should be performed to treat osteosarcoma, and resection of tumor tissue is an important step in osteosarcoma treatment. Preoperative prophase chemical or radiation therapy and postoperative consolidation chemical or radiation therapy are very important for creating operation conditions, inhibiting tumor metastasis and improving the survival rate of patients. In addition, recent therapeutic approaches such as immunotherapy and molecular targeted therapy have been advanced to some extent in recent years, but the therapeutic effect is not yet established.

Accordingly, there is a continuing unmet need in the art for the development of new drugs effective in the treatment of osteosarcoma.

Disclosure of Invention

The invention aims to enrich clinical medication choices and provide a pharmaceutical composition for treating infantile osteosarcoma.

The present inventors have unexpectedly found that 2-methyl-2- (4- {5- [3- (methylsulfonyl) phenyl ] -1H-pyrrolo [2,3-b ] pyridin-3-yl } phenyl) propanoic acid (hereinafter referred to as compound I) has an excellent effect of inhibiting the growth of osteosarcoma cells, and thus can be expected to be particularly useful in the treatment of pediatric osteosarcoma

Compound I is a known compound, as originally disclosed in International patent application WO2006/063167A1 (which is incorporated herein by reference in its entirety) to Smithkline Beecham Corporation. WO2006/063167A1 relates generally to 1H-pyrrolo [2,3-b ] inhibitors useful as serum and glucocorticoid-regulated kinase-1 (SGK-1) inhibitors]Pyridine compounds, pharmaceutical compositions containing such compounds, and methods of using such compounds and their pharmaceutical compositions to treat SGK-1 mediated diseases, particularly certain forms of renal disease, congestive heart failure. In the specification of this patent document, compound I is disclosed as example 203, which describes the preparation method, physical properties and¹H-NMR data. However, this patent document does not mention that the compound has an effect of inhibiting the growth of osteosarcoma cells, which constitutes an unexpected finding of the present invention.

To this end, the present invention provides a pharmaceutical composition for the treatment of pediatric osteosarcoma, said pharmaceutical composition comprising a) Compound I or a pharmaceutically acceptable salt thereof and b) a pharmaceutically acceptable excipient.

In one aspect, the pharmaceutical composition comprises 0.01 to 90%, preferably 0.1 to 60%, more preferably 1 to 50% of compound I or a pharmaceutically acceptable salt thereof, based on the total weight of the pharmaceutical composition.

The compound I or a pharmaceutically acceptable salt thereof may be used in combination with other active ingredients for the treatment of pediatric osteosarcoma. According to this aspect, the pharmaceutical composition may further comprise other pharmaceutically active ingredients for the treatment of pediatric osteosarcoma selected from, but not limited to, one or more of doxorubicin, cisplatin, methotrexate, ifosfamide or pharmaceutically acceptable salts thereof. The other pharmaceutically active ingredient may also be formulated separately in another pharmaceutical composition and administered simultaneously or sequentially with the pharmaceutical composition of the invention.

The term "pharmaceutically acceptable salt" is used in the sense as is commonly understood in the art. For example, preferred pharmaceutically acceptable salts in the present invention may include base addition salts, e.g., pharmaceutically acceptable base addition salts of compound I with alkali metals, alkaline earth metals, ammonia or organic amines, such as, but not limited to, lithium, sodium, potassium, magnesium, calcium, ammonium, ethylenediamine salts.

"pharmaceutically acceptable excipient" is used in the sense as commonly understood in the art. The present invention does not particularly limit the kind of the pharmaceutically acceptable excipient, so long as it is compatible with the active ingredient used and suitable for the intended administration route. For example, when the active ingredient of the present invention is intended to be prepared into tablets, fillers such as lactose, microcrystalline cellulose, mannitol; binders such as hypromellose, povidone, starch slurry; disintegrants such as croscarmellose sodium, sodium carboxymethyl starch, hydroxypropyl starch; lubricants such as talc, magnesium stearate, aerosil; and optionally coloring agents, sweeteners, coating materials and the like. For another example, when the active ingredient of the present invention is intended to be prepared into an injection, various sterile excipients known in the art, such as water for injection; oils for injection such as castor oil, soybean oil, peanut oil; and other solvents for injection such as propylene glycol, glycerol, polyethylene glycol, ethyl oleate; and optionally emulsifying agents, suspending agents, solubilizing agents, buffering agents, bacteriostats, antioxidants, isotonicity adjusting agents and the like.

The pharmaceutical compositions of the present invention may be administered by any suitable route of administration. Preferably, the pharmaceutical composition according to the invention is administered by oral or parenteral route. It will be appreciated that the appropriate route of administration will depend upon such factors as the sex, age, weight, general medical condition of the patient to be treated, the severity of the disease being treated and the like, and will be determined by the attending clinician based upon the diagnostic results and clinical experience.

The invention also provides application of the compound I or the pharmaceutically acceptable salt thereof in preparing a pharmaceutical composition for treating infantile osteosarcoma.

In order that the nature and spirit of the present invention may be further understood, preferred embodiments of the present invention and the effects thereof will be described below with reference to specific examples. It is to be understood, however, that such description is merely illustrative of the features and advantages of the present invention, and is not intended to limit the scope of the appended claims in any way.

Detailed Description

Examples

Tablet formulation

The preparation method comprises the following steps: compound I, calcium sulfate dihydrate and lactose were mixed well and granulated with an appropriate amount of 5% hypromellose aqueous solution. Sieving the wet granules with a 80 mesh sieve, drying, and mixing with starch, talc powder and magnesium stearate; sieving with 40 mesh sieve, and making into tablet, and making into 200 tablets.

Experimental example 1 evaluation of in vitro proliferation inhibitory Effect of Compound I on osteosarcoma cells

The purpose of this experiment was to investigate the inhibitory effect of compound I on the in vitro cell proliferation of two osteosarcoma cell lines, human osteosarcoma cell line MG63 and rat osteosarcoma cell line UMR 106.

1. Experimental methods

MG63 and UMR106 cells grown in DMEM medium containing 10% fetal bovine serum in exponential growth phase were grown at 4.5X 10³The cells were seeded at a density of one cell/well in 96-well culture plates (90. mu.l per well volume) and then returned to the incubator for further incubation at 37 ℃ in 5% carbon dioxide for 24 hours. The 96-well plate was then removed from the incubator and the drugs were applied according to the following dosing schedule (10 multiple wells per schedule):

blank control group: to each well was added 10. mu.L of the above DMEM medium.

Positive control group: to each well was added 10. mu.L of the above DMEM medium containing 0.5. mu.g/mL doxorubicin hydrochloride.

Compound I low concentration group: to each well was added 10. mu.L of the above DMEM medium containing 0.1. mu.g/mL of Compound I.

Concentration group in compound I: to each well was added 10. mu.L of the above DMEM medium containing 0.5. mu.g/mL of Compound I.

Compound I high concentration group: to each well was added 10. mu.L of the above DMEM medium containing 1.0. mu.g/mL of Compound I.

Then, the 96-well plate was returned to the incubator to continue the incubation for 48 hours. After completion of the incubation, 20. mu.L of MTT (5 mg/mL PBS) was added to each well of the 96-well plate in the dark, and the plate was returned to the incubator and incubated at 37 ℃ under 5% carbon dioxide for 3 hours. Then, all the medium was aspirated, 120. mu.L of DMSO was added, the mixture was shaken in a water bath at 37 ℃ for 30min, and after the resulting purple crystals were completely dissolved, the absorbance value (OD) at a wavelength of 570nm was measured with a microplate reader₅₇₀). The inhibition of in vitro cell proliferation of MG63 and UMR106 cells by each dosing regimen was evaluated using the inhibition ratio (%) calculated as follows: inhibition rate (%) [1- (OD of administration group)₅₇₀value/OD of blank control₅₇₀Value)]×100％。

2. Results of the experiment

The results of this experiment are shown in tables 1-2 below.

TABLE 1 inhibition ratio of Compound I on in vitro cell proliferation of human osteosarcoma cell line MG63 (%)

Group of	Inhibition ratio (%)
		Blank control group	0
Positive control group	23.9±6.5
		Compound I Low dose group	15.7±6.1
Compound I Medium dose group	38.3±8.9*
		High dose group of Compound I	56.2±9.6*

TABLE 2 inhibition of in vitro cell proliferation of compound I on rat osteosarcoma cell line UMR106 (%)

Note: the results are expressed as mean ± standard deviation, indicating p <0.05 compared to the positive control group.

3. Discussion of the related Art

The above results indicate that compound I has certain cell proliferation inhibitory effect on both osteosarcoma cell lines tested, human osteosarcoma cell line MG63 and rat osteosarcoma cell line UMR106, and shows certain dose dependence. Moreover, under the same concentration condition, the proliferation inhibition effect of the compound I on the two osteosarcoma cell lines is even higher than that of a positive control drug, namely doxorubicin hydrochloride.

Experimental example 2 in vivo antitumor Effect of Compound I on osteosarcoma-bearing mice

The purpose of this experiment was to investigate the in vivo therapeutic effect of compound I on UMR106 tumor-bearing mice.

1. Experimental methods

Male C57BL/6 mice, 6-8 weeks old, were used in this experiment. Animals were acclimated for two days at 23 ℃ room temperature, 50% relative humidity and 12 hours light/12 hours dark conditions. Animals can eat and drink water freely.

On the day of the experiment, UMR106 cells were cultured at 8X 10⁵The dose of/100. mu.L/mouse was inoculated subcutaneously to mice with no statistical difference in body weight. When the tumor grows to about 100mm³(tumor volume was estimated according to the following formula: V ═ 0.52X length X width²Unit mm³) At the time, tumor-bearing mice were randomly divided into 4 groups of 8 mice each, and each experimental group was administered according to the following dosing schedule:

blank control group: normal food and drinking water were given for 28 days.

Compound I low dose group: in addition to normal food and drinking water, mice were also gavaged with compound I between 9 and 10 am daily at a dose of 10mg/kg body weight/day for 28 days.

Dose groups in compound I: in addition to normal food and drinking water, mice were also gavaged with compound I between 9 and 10 am daily at a dose of 50mg/kg body weight/day for 28 days.

Compound I high dose group: in addition to normal food and drinking water, mice were also gavaged between 9 and 10 am daily with compound I at a dose of 100mg/kg body weight/day for 28 days.

Mice were observed daily for activity status and tumor volume growth. After dosing was complete, mice were sacrificed by cervical dislocation, tumors were removed, tumor volume was measured and weighed.

2. Results of the experiment

The results of this experiment are shown in table 3.

TABLE 3 Effect of Compound I on tumor volume and weight in UMR106 tumor-bearing mice

Note: the results are expressed as mean ± standard deviation, indicating p <0.05 compared to the blank control group.

3. Discussion of the related Art

The experimental results show that the compound I has obvious therapeutic action on the tumor of the UMR106 tumor-bearing mouse and is consistent with the results of in vitro experiments. Compared with the blank control group, the mice of the low, medium and high dose groups of the compound I have obviously reduced tumor growth, and the volume and the weight of the tumor are obviously reduced.

In summary, the above experimental results fully confirm that the compound I can be used for preparing a pharmaceutical composition for treating pediatric osteosarcoma, and provide more clinical medication options for the means for treating pediatric osteosarcoma in the field.

The foregoing is only a preferred embodiment of the present invention. It should be noted that, for those skilled in the art, without departing from the spirit and principle of the present invention, several improvements, modifications, equivalents and the like can be made, and these improvements, modifications, equivalents and the like also should be regarded as falling within the protection scope of the present invention.

Claims

Use of 2-methyl-2- (4- {5- [3- (methylsulfonyl) phenyl ] -1H-pyrrolo [2,3-b ] pyridin-3-yl } phenyl) propanoic acid or a pharmaceutically acceptable salt thereof in the manufacture of a pharmaceutical composition for the treatment of pediatric osteosarcoma.