CN106860458A

CN106860458A - Application, method and pharmaceutical composition of the quinine class compound in tumour is treated

Info

Publication number: CN106860458A
Application number: CN201510922532.5A
Authority: CN
Inventors: 邹方霖; 卢星; 邹礼常; 王建霞
Original assignee: Chengdu Kuachang Science and Technology Co Ltd
Current assignee: Chengdu Kuachang Science and Technology Co Ltd
Priority date: 2015-12-11
Filing date: 2015-12-11
Publication date: 2017-06-20

Abstract

The application in tumour is treated and method the present invention relates to quinine class compound, and the pharmaceutical composition comprising quinine class compound.

Description

Application, method and pharmaceutical composition of the quinine class compound in tumour is treated

Technical field

The application in tumour is treated and method the present invention relates to quinine class compound, and comprising quinine class The pharmaceutical composition of compound.

Background technology

Many quinine class compounds are antiparasitic, especially anti-malarial agents.Quinine class compound antimalarial it Outer application, is much carried out with composition forms.This based composition is once applied to treat various diseases, Such as hemorrhoid, rhinitis, hemangioma, facial paralysis etc..It is generally believed that other components (such as crow drawings in composition It is smooth) not only some drug effects also can in the treatment be played with hydrotropy.Thus, include quinine for specified disease The pharmaceutical composition of class compound is always among research.For example, Chinese patent 87104056 discloses one kind Six components compositions " injection for whole prolapse haemorrhoids " containing a quinin hydrochloride and quinine dihydrochloride, for remove hemorrhoid and Anal fissure, rectal polyp and the papilla fiber histioma associated with hemorrhoid.But in the composition, quinine class Compound total concentration is more than 0.47M, and contains urethane.Chinese patent 200910103187.7 provides one Nine components compositions are planted to be used to treat hemorrhoid and tumour.The concentration very little (being less than 0.01M) of quinine in said composition, And contain urethane.Because the side effect of urethane is stronger, Chinese Government has limited or even has forbidden this kind of answering With.Chinese patent 96110932 discloses a kind of composition comprising a quinin hydrochloride and Anodynine, its use In Treatment of Hyperthyroidism.However, Anodynine side effect is very strong, in developed country, oneself is eliminated.

Possible application of the quinine class compound in terms of anti-knurl once received very many concerns.In fact, In in vitro test with tumour cell as model, some quinine class compounds and conventional antitumor medicine (such as 5- fluorine Pyrimidine, adriamycin, taxol etc.) cell 503nhibiting concentration (IC₅₀) all every liter of micromole's (μM ol/L) In level.According to common understanding, a concentration dependent for the Antineoplastic effect of medicine is in IC₅₀Afterwards with concentration Increase and reduce.Conventional antitumor medicine is generally with than IC₅₀The concentration of several times is higher by (still in every liter of scope of micromole It is interior) when being injected in knurl body, just show anti-knurl validity.Even if however, we are quinine class compound (example Such as quinine dihydrochloride) injection concentration improve about 500 times (0.025M), effective anti-knurl effect is also not observed Really (tumour inhibiting rate is less than 40%), does not show the drug effect more considerably higher than existing antitumor medicine.In the prior art, The tumour inhibiting rate of other quinine class medicines is not also high.In the last few years, they were just no longer regarded as one kind by people competition The antitumor medicine of power, and be treated as anti-knurl activity increase agent and studied.For example, in No. 201310459568.5 Quinine and antitumor medicine (vincristine class medicine) are exactly used to reduce the multiple medicine of cell by state's patent application Drug resistance is so that antitumor medicine reaches preferable drug effect.

The treatment of current tumour, the mainly chemotherapy of operation, radiation cure and other medicines.Operation is to body Body is damaged, and the injury of the gland (such as mammary gland and thyroid gland) attractive in appearance on some influences is frustrating, Also it is made to become a kind of painful choice.The side effect of radiation cure is also clearly.Existing chemotherapy is substantially It is systemic administration, there is systemic side effects risk very high.For example, existing antitumor medicine is in interior killing on a large scale While cancer cell, a large amount of normal cells are also killed, immune system is seriously damaged, and curative effect is also simultaneously It is unsatisfactory.

The content of the invention

Goal of the invention of the invention be to provide it is a kind of compared with prior art more effectively smelting treat tumour medicine Composition and treatment method.

According to an aspect of the present invention, it provides a kind of pharmaceutical composition for treating tumour, and it is included Quinine class compound and pharmaceutically acceptable excipient, and be the formulation for being suitable for being administered through knurl area, wherein When being administered through knurl area, the concentration of quinine class compound is 0.05-0.5M in described pharmaceutical composition, preferably 0.15-0.4M, more preferably 0.2-0.3M.

According to another aspect of the present invention, it provides application of the quinine class compound in tumour is treated, its Described in quinine class compound be included in pharmaceutical composition, described pharmaceutical composition is comprising pharmaceutically acceptable Excipient and be the formulation for being suitable for being administered through knurl area, wherein when knurl area is administered, Kui in described pharmaceutical composition The concentration of peaceful class compound is 0.05-0.5M, more preferably preferably 0.15-0.4M, 0.2-0.3M.

According to a further aspect of the invention, it provides a kind of method for treating tumour, and it is included to there is this to need The individual knurl area administration wanted pharmaceutical composition of acceptable excipient comprising quinine class compound and pharmaceutically, Wherein described pharmaceutical composition is the formulation for being suitable for being administered through knurl area, and when the knurl area is administered, the medicine In compositions the concentration of quinine class compound be 0.05-0.5M, preferably 0.15-0.4M, more preferably 0.2-0.3M。

Within the scope of the invention, when knurl area is administered, the administered volume of described pharmaceutical composition is not less than desire 70%, preferably not less than the 110% of the tumor mass volume for the treatment of.

Within the scope of the invention, the pharmaceutically acceptable excipient includes water, water is miscible has Machine solvent or their mixture, wherein the miscible organic solvent of the water includes one below kind or various： Ethanol, propane diols, PEG, isopropanol.Pharmaceutical composition of the invention does not include such as urethane and peace For being prohibited because side effect is relatively strong than woods etc. or limit the drug ingedient for using.

Within the scope of the invention, it is described in addition to quinine class compound and pharmaceutically acceptable excipient Composition can also further include anodyne, other antineoplastics and/or synergist.

Within the scope of the invention, the tumour includes malignant tumour and benign tumour, the malignant tumour example Breast cancer, liver cancer, cancer of pancreas, thyroid cancer, prostate cancer, lung cancer, head and neck cancer, colon cancer, nose in this way The various malignant entity tumors such as pharynx cancer, and the benign tumour include various benign entity tumors, such as mastadenoma, Hepatoma, thyroid adenoma, prostate tumor etc..

Embodiment of the invention has advantages below compared with prior art：With it is existing operation and put Beta ray therapy is compared, and shows that same efficient more friendly " treatment-patient body " relation (is disturbed to body It is smaller)；Compared with existing systemic administration is treated, can be carried to conspicuousness under the toxic and side effect for minimizing Curative effect high；Compared with existing local application, especially curing agent (such as absolute ethyl alcohol) are treated, with lesion Compatibility between surrounding tissue is higher, and better efficacy.

The present invention is described in more detail below with reference to accompanying drawing.

Brief description of the drawings

Fig. 1 is the injection concentration-tumour inhibiting rate relation curve of quinine dihydrochloride injection, and wherein abscissa X is knurl Area's injection concentration (4 × 10^XμM) index (X), and ordinate Y is the tumour inhibiting rate (Y%) measured in zoopery.

Specific embodiment

Within the scope of this invention, term " quinine class compound " refers to quinine (Quinine), quinine isomery Body and their derivative.Quinine isomers for example can be quinindium, cinchonine and cinchonidine.It is described to spread out Biology includes acid-addition salts, such as inorganic acid addition salt and organic acid addition salt.Inorganic acid addition salt for example can be with Be quinine or quinine isomers halogen acid salt (such as hydrochloride, hydrobromate, hydriodate, dihydrochloride, Dihydrobromide, two hydriodates), disulfate, sulfate, phosphate.Organic acid addition salt for example may be used Being bicarbonate or carbonate, ethyl carbonate salt, formates, acetate, the Chinese holly of quinine or quinine isomers Edge hydrochlorate or tannate., according to the invention it is preferred to quinine class compound be, for example, quinine, a quinin hydrochloride And quinine dihydrochloride.

Within the scope of this invention, pharmaceutically acceptable excipient (or solvent) can be water, water can Misci-ble organic solvents or the vehicle system comprising water and the miscible organic solvent of water.Water is miscible to be had Machine solvent includes one below kind or various：Ethanol, propane diols, polyethylene glycol (PEG), isopropanol.Its Middle PEG can optionally employ PEG, such as PEG200, PEG300 and PEG400 of the molecular weight less than 600 Deng.

The anodyne can be any appropriate person well known by persons skilled in the art, to mitigate the pain of patient Sense, such as phenmethylol, procaine hydrochloride, anesin, hydrochloric acid benefit card etc..

Described other antineoplastics can be any appropriate person well known by persons skilled in the art, further to increase Powerful antitumor activity, for example including adriamycin, 5-FU, gemcitabine, cis-platinum, chloroquine, primaquine, Qinghaosu and its derivative etc..

Within the scope of the invention, in addition to quinine class compound and pharmaceutically acceptable excipient, described group Compound can also further include synergist.The synergist can be any conjunction well known by persons skilled in the art Suitable person, to further enhance treatment tumor effect, such as including curing agent and activity increase agent.

The curing agent can be any appropriate person well known by persons skilled in the art, add it to promote tumour Necrosis, reduce and fibrosis, it for example can be Lauromacrogol, sodium morrhuate, EO, poly- many Card alcohol and bleomycin A5 etc..

In the present invention, term " activity increase agent " refers to that activity is compared smaller or even there is no (such as in animal 40%) tumour inhibiting rate in experiment is less than but can be used to increase the material of other medicines anti-tumor activity, including immune Adjuvant.The immunologic adjuvant can be any appropriate person well known by persons skilled in the art, add it to promote to suffer from The activity of the related antigen for being beneficial to treat tumour in lesion, it for example can be inorganic adjuvant, such as hydroxide Aluminium, alum etc.；Microorganism and its product such as mycobacteria (tubercle bacillus, BCG vaccine), bacillus pumilis, hundred Day coughs bacillus, endotoxin, bacterial extract (muramyl dipeptide) etc.；Synthetic adjuvant, such as artificial synthesized is double Chain polynucleotide (double-strand polyadenylic acid, uridylic acid), levamisol, isoprinosine etc.；Finish, such as Fei Shi adjuvants, adjuvant 65, mineral oil, vegetable oil etc.；Immunostimulant (BCG vaccine, rod-like stem Bacterium, endotoxin, trehalose, thymic peptide, OK432 etc.)；Cell factor, such as interferon, interleukins, TNF, TGF, colony stimulating factor, chemotactic factor (CF), thymosin extrasin etc.；Heterogenetic antigen, Such as inactivate streptococcus, HRBC's membranous antigen, tumor infiltrating lymphocyte, tumor vaccine.

Term " administration of knurl area " refers to that medicine is administered directly to including knurl body and Liu Zhouliu areas.Knurl area is administered Operating can be realized by all multi-methods, such as injection, conduit introducing, arterial perfusion etc..When knurl area is administered, Pharmaceutical composition of the invention is the formulation for being suitable for the administration of knurl area, for example, can be injection etc., and Kui Concentration of the peaceful class compound in the pharmaceutical composition that the knurl area is administered is 0.05-0.5M (Mol/L), preferably 0.15-0.4M, more preferably 0.2-0.3M.In addition, when knurl area is administered, the administration of described pharmaceutical composition Volume is not less than the 70% of the tumor mass volume for being intended to process, 110% is preferably not less than, so that comprising quinine class The pharmaceutical composition of compound can infiltrate knurl body and its peripheral region, realize antitumor action higher.If treated The volume for processing tumour is sufficiently small, then its volume is the above-mentioned volume for being intended to process.When gross tumor volume is larger, Then need to be allocated as the medication in batches of multiple parts, then the part volume is the above-mentioned volume for being intended to process.

For the application of the present invention and the pharmaceutical composition of method Zhong Jingliu areas administration, by taking injection as an example, can Prepare in accordance with the following methods.

Method one：Solid phase quinine class compound for required concentration in water-soluble concentration range, such as concentration is small In the quinine dihydrochloride of 0.35M, and water-soluble strong anodyne, other antineoplastics being optionally present And/or anti-knurl synergist, obtain corresponding injection by they are directly added into water for injection.

Method two：Solid phase quinine class compound for required concentration outside water-soluble concentration range, such as concentration A quinin hydrochloride more than 0.15M, and the poorly water-soluble being optionally present anodyne, other antineoplastics Thing and/or anti-knurl synergist, then first prepare required vehicle system (such as 9% ethanol solution), then will be upper State during solid matter adds the vehicle system, you can obtain the injection of the quinine class compound of concentration comprising needed for.

Method three：When the anodyne for being insoluble in water, other antineoplastics and/or anti-knurl synergist need to be added, Then can for example prepare by the following method：

A. quinine class compound water solution, or the solution for preparing required vehicle system and quinine class compound are prepared；

B. vehicle system needed for preparing and anodyne, other antineoplastics and/or the anti-knurl synergy for being insoluble in water The solution of agent；

C. the solution that a and b is prepared is mixed.

By above-mentioned these methods, the various injections comprising quinine class compound can be prepared.For example, different notes The change penetrated in agent includes：Containing different quinine class compounds, the quinine class compound containing various concentrations, containing not With the miscible organic solvent of water, the miscible organic solvent of water containing various concentrations, containing variety classes and dense The anodyne of degree, other antineoplastics and/or synergist, etc..But change anyway, quinine class Total concentration of the compound in the pharmaceutical composition that knurl area is administered should remain 0.05-0.5M (Mol/L), preferably 0.15-0.4M, more preferably 0.2-0.3M.

If it does, the concentration of anodyne is 0-3% (weight)；The concentration of other tumour medicines is 0-0.15M； The concentration of synergist is 0-3% (weight).

Certainly, the injection concentration comprising quinine class compound (such as quinine dihydrochloride) needed for the present invention also may be used Obtained by diluting high-concentration quinine class compound injection liquid (such as containing the parenteral solution of 25W/V% quinine dihydrochlorides) .

Within the scope of the invention, the pharmaceutical composition comprising quinine class compound can also before knurl area is administered It is the form of solid powder, is restored with corresponding solvent before administration.

Within the scope of the invention, it is to increase effective acting time of the quinine class compound at administration, improve it Bioavilability, it can for example mix micro particles, nano-particle, the micella as slow-released carrier or matrix In situ-gel matrix.

Embodiment

By specific examples below, the present invention is further illustrated, but not as limitation of the present invention.

In the examples below, unless otherwise indicated, subcutaneous transplantation knurl animal experiment is carried out as follows.

Animal：SPF grades of nude mice (BALB/C-Nude), all females of sex, animal about 6~8 during packet Week old, body weight is 17.5-20.5g.

Animal inoculation pvaccination and packet：The tested tumour cell of in vitro culture, collects the tumour cell of exponential phase, will The tumor cell inoculation of required number is in the subcutaneous of nude mice.Tested tumour cell includes：Human liver cancer cell (HepG2), human breast cancer cell (MDA-MB231), human lung carcinoma cell (A549), human thyroid cancer cell (SW579), Human Prostate Cancer Cells (LNCaP/AR), human pancreatic cancer cell (PANC-1), people's colon Cancer cell (COLO205), people's head ＆ neck cancer cell (Fuda), KB cell (CNE1), human gastric cancer (BGC823), oophoroma (PA1) etc..Research method and result in embodiments of the invention are also applied for quinine Class compound has other tumour cells of anti-tumor activity to it.Treat that tumour is long to 50-100mm³When, use The softwares of PEMS 3.2 (establishment of West China HSPH of Sichuan University) carry out random district's groups to animal subject, respectively It is negative control group, positive controls and several drug research groups, every group 8.

Administration：The packet same day starts administration, and injection volume is determined by dosage and concentration.Administration number of times is 6 times, is given Medicine frequency is per once two days.The injected material of negative control group is corresponding solvent.The sun of positive controls injection Property tester be selected from above-mentioned existing antitumor medicine, administering mode and dosage carry out (example by the convention of selected medicine Such as tail vein injection).Drug research group needs to inject the research medicine of variety classes and concentration according to experiment, removes It is otherwise noted and is the injection of knurl area.

Observation, measurement and analysis：

1) general state：Observation 1 time daily, self administration of medication starts to experiment the 28th day, observation index or content Including but not limited to animal is administered situations such as part, appearance sign, general behavior activity, the state of mind, death And other Novel presentations.

2) body weight：Quarantine determines 1 time, is determined 1 time before administration and administration phase is determined weekly 2 times.

3) food ration：Administration phase is determined weekly 1 time.

4) relative tumor growth inhibiting rate：Measured in the 1st, 3,5,7,11,16,21,27 days after administration Transplantable tumor size.

Gross tumor volume computing formula is as follows：TV=l/2 × a × b², a represents length of tumor in formula, and b represents swollen Knurl width.

Relative tumour volume computing formula is as follows：RTV=V_t/V₀, V in formula₀It is the packet administration same day (i.e. the One day) measurement obtained by gross tumor volume, V_tGross tumor volume during to measure each time.

Relative tumor proliferation rate computing formula is as follows：T/C (%)=T_RTV/C_RTV× 100, T in formula_RTVIt is the positive The RTV of control group or drug research group, C_RTVIt is the RTV of negative control group.

The evaluating drug effect standard of drug research group is：T/C (%)>40 for inactive, T/C (%)≤40 (but>15%) And Analysis of variance P compared with negative control group<0.05 is active (without preferred agents activity), T/C (%)≤15 and Analysis of variance P compared with negative control group<0.05 is have preferred agents activity.

5) inhibition rate of tumor growth (tumour inhibiting rate) is determined：System is carried out after being administered in first time within the 28th day to become celestial, Knurl body is carried out to weigh.Tumour inhibiting rate computing formula is as follows：Tumour inhibiting rate %=(TW-CW)/CW × 100%, wherein TW is the average knurl weight of positive controls or drug research group；CW is the average knurl weight of negative control group.

The evaluating drug effect standard of drug research group is：Tumour inhibiting rate<40% for inactive, tumour inhibiting rate >=40% (but <And Analysis of variance P compared with negative control group 85%)<0.05 for active, tumour inhibiting rate >=85% and through variance Analysis P compared with negative control group<0.05 is have preferred agents activity.

In the examples below, zoopery uses duplicate measurements variance analysis (Repeated Measures ANOVA) group differences respectively to index mean carry out statistical test.Group differences are statistically significant When (P≤0.05), each group is compared with negative control group difference using least significant difference method.It is quantitative Index is described using mean ± standard error (X ± SEM).When LEVENE homogeneitys test of variance point out variance not When neat (P≤0.05), group difference is compared using Mann-Whitney U rank tests (M-W methods).Institute Some statistical analyses, complete under the softwares of SPSS for Windows 13.0.

Embodiment 1：The preparation of quinine class compound injection agent

Table 1

Remarks:+ to refer in addition to the composition in table remaining be full water for injection

- refer to being free of.

In table 1, the preparation method of A2 is as follows：At room temperature, 0.2ml phenmethylols are added into 8.5ml injections Dissolved with water, dissolved during 758mg quinine dihydrochloride dry powder then is added into the solution, add injection Water makes cumulative volume reach 10ml, it is well mixed after be packed as 2ml/ bottle standby, just prepared injection A2.

The preparation method of A3 is as follows：At room temperature, will be molten in 0.2ml phenmethylols addition 8.0ml waters for injection Solution, dissolves during 758mg quinine dihydrochloride dry powder then is added into the solution, adds 1000mg polidocanols Dissolving, being eventually adding water for injection makes cumulative volume reach 10ml, it is well mixed after be packed as 2ml/ bottles it is standby, Just injection A3 is obtained.

The preparation method of A4 is as follows：At room temperature, will be molten in 0.2ml phenmethylols addition 8.0ml waters for injection Solution, dissolves during 379mg quinine dihydrochloride dry powder then is added into the solution, adds 516mg chloroquine diphosphates Dissolving, being eventually adding water for injection makes cumulative volume reach 10ml, it is well mixed after be packed as 2ml/ bottles it is standby, Just injection A4 is obtained.

The preparation method of B2 is as follows：At room temperature, by 0.9ml absolute ethyl alcohols addition 7.5ml waters for injection Mix, then 0.2ml phenmethylols are added thereto dissolving, then add this molten in the quinin hydrochloride dry powder of 595mg mono- Dissolved in liquid, adding water for injection makes cumulative volume reach 10ml, it is well mixed after be packed as 2ml/ bottles it is standby, Just injection B2 is obtained.

The preparation method of B3 is as follows：At room temperature, by 0.9ml absolute ethyl alcohols addition 7.5ml waters for injection Mix, then 0.2ml phenmethylols are added thereto dissolving, then add this molten in the quinin hydrochloride dry powder of 595mg mono- Dissolved in liquid, add the dissolving of 1g polidocanols, being eventually adding water for injection makes cumulative volume reach 10ml, mixes Close it is uniform after be packed as 2ml/ bottle standby, just prepared injection B3.

NM A1, A5, A6 and B1, B4 and C1 are according to method preparation similar to the above .

The injection concentration of the required quinine class compound (such as quinine dihydrochloride) in the present invention, also can be by dilute High concentration quinine class compound (such as quinine dihydrochloride of W/V percentages 25%) parenteral solution is released to obtain.

Embodiment 2：The optimum condition research of application

In the experiment of anti-knurl, positive control is usually existing clinical medicine (such as cis-platinum, adriamycin, mitogen Mycin, gemcitabine, 5-FU etc.).Their clinical meaning is hypotoxicity high activity, such as dynamic Hypotoxicity and the tumour inhibiting rate up to 75% are shown in thing experiment.Term " anti-tumor activity ", it is sometimes again referred to as active, Refer to that there is certain anti-homophylic property of knurl compared with existing clinical medicine.During a kind of material as tested in vitro (IC is abbreviated as with relatively low 503nhibiting concentration₅₀, 50 μM are less than in generally suppressing reaction at 72 hours) or Show that (abbreviation tumour inhibiting rate typically larger than 40% but is less than for inhibition rate of tumor growth higher in zoopery 75%), just it is considered as having similarity with existing clinical medicine, so as to show anti-tumor activity, the material Also just it is referred to as active material.Term " reactive conditions " refers to that the something that makes that researcher provides shows that it resists The condition of tumor activity.For example, making the tumour inhibiting rate of quinine class compound bring up to 75% or so from 35% or so Condition (application method, dosage etc.).

Term " preferably anti-tumor activity ", it is sometimes again referred to as preferred active, refer to have compared with existing clinical medicine There is the property of more hypotoxicity and considerably higher anti-tumor activity.For example, it is preferable to activity can refer in zoopery with Less side reaction cost (non-toxic only slight side effect) shows significantly greater tumour inhibiting rate (such as 85% More than).Term " optimum condition " refers to that the something that can make that researcher provides shows that its preferably anti-knurl is lived The condition of property.Term " preferred substance " refers to not only give anti-tumor activity condition and give optimum condition Material.

The optimum condition of application of the quinine class compound of the invention in oncotherapy, anti-knurl is lived in vitro first Property experiment and transplantable tumor animal model on studied.

1) anti-tumor activities

In the present invention, medium effective concentration (is abbreviated as IC₅₀) when being that cell survival rate reduces 50% in experiment in vitro Drug concentration.11 plants of man―machine systems are selected in this experiment, and medicine is determined using tetrazolium MTT reducing process The antitumor action of thing.Experiment sets positive and negative control group and seminar.Negative control group adds isometric life Reason salt solution, positive controls are the conventional chemotherapeutic drugs 5-FU of various concentrations (in μM ol/l levels).Grind The quinine class compound for studying carefully the various concentrations that group (B, C, D) group is respectively diluted with higher concentration mother liquor is molten Liquid (in μM ol/l levels).These mother liquors be respectively embodiment 1 preparation containing the quinin hydrochlorides of 0.25M mono-, The injection of 0.25M quinine dihydrochlorides and 0.15M quinines, wherein containing 2% anodyne phenmethylol.

In experiment, from the attached tumor cells of exponential phase, after being digested with pancreatin, with containing 10% calf The RPMI l640 culture mediums of serum are made into the suspension of required cell concentration, are seeded in 96 well culture plates, often Hole is inoculated with 200 μ l, then at 37 DEG C and 5%CO₂Middle culture 24h.What seminar and positive controls renewed Culture medium containing various concentrations sample, negative control group then changes the culture medium containing isometric physiological saline, often Group sets 5 parallel holes, makes final volume for 200ul/ holes, cultivates 48 hours.Then abandoning supernatant, per hole The serum free medium containing 0.2mg/ml MTT of 200 μ l Fresh is added, continues to cultivate at 37 DEG C 4h.It is careful to abandon supernatant, and 200 μ l DMSO are added, after being mixed with miniature ultrasonic oscillator, in ELIASA On with tested wavelength as 570nm, reference wavelength be 450nm determine OD value.It is calculated as follows medicine pair The inhibiting rate of growth of tumour cell：Growth of tumour cell inhibiting rate %=(1-OD experiment/OD controls) × 100%

Dose-effect curve can obtain to inhibition rate of tumor cell mapping with same sample various concentrations, then adopt Medium effective concentration (IC is calculated with Logit methods₅₀Value).Usual IC₅₀Concentration be less than 100 μM when, then judge Sample has lethal effect to tumour cell in vitro.Table 2 is test composition to testing the IC of tumour cell₅₀, Concentration value is in terms of the quinine class compound concentration in composition.Generally speaking, they all μM concentration level, Significant difference is not seen statistically.And the IC of positive control₅₀Then between 0.01-0.5 μM.

Table 2

By Try tumour cells
				Human breast carcinoma MDA-MB231	62.2	56.5	73.5
Human lung adenocarcinoma A549	48.9	51.4	62.8
				Oophoroma PA1	18.7	13.4	29.2
Stomach cancer BGC823	72.8	66.8	49.5
				Colorectal cancer LOVO	-	103	-
Human liver cancer HepG2	31.9	32.6	42.1
				Human colon carcinoma LS174T	78.4	62.9	72.9
Human prostata cancer LNCaP/AR	32.9	28.9	36.4
				People's head and neck cancer Fuda	62.1	51.6	45.8
Human thyroid cancer cell SW579	35.6	46.8	72.3
				Human pancreatic cancer cell PANC-1	45.7	31.7	42.9

Further test and also demonstrate, the organic solvent in the concentration range of introducing and analgesic in the present composition Agent has no negative effect to the anti-tumor activity of quinine dihydrochloride.

2) application method

Experimental animal is lotus HCC nude mice, is divided into negative control group, positive controls and drug research group. Negative control group injecting normal saline, the conventional antitumor medicine 5-FU of positive controls injection, drug research group Injection quinine dihydrochloride solution.When tail vein injection is small, in, heavy dose of concentration be two hydrochloric acid of 0.025M During the quinine aqueous solution, although we have also made some technical improvement, eligible result still with the knot of prior art Fruit is consistent, and tumour inhibiting rate is less than 30%.And reach 75% as the tumour inhibiting rate of the 5-FU of positive control.

When we prepare to dilute standby 0.25M quinine dihydrochlorides (containing 2% anodyne) direct injection a kind of When on to animal knurl body, unexpected result is but occurred in that.Although target area administration as you know may improve The efficiency (for example making tumour inhibiting rate increase, generally to improve 10-30%) of systemic administration, and our achievement in research But drug effect mysterious height (can improve more than 90%) has been brought up to.Conversely, said composition passes through tail During intravenous injection, tumour inhibiting rate is again below 30%.

The administration of knurl area depends not only on drug effect, also depends on its security.Therefore, we compare 0.25M bis- The animal tail vein injection of quinin hydrochloride injection and the maximum tolerated dose (Maximum of Lump body injection Tolerated dose, MTO), method is as follows：

Experimental animal be lotus HCC nude mice, be grouped at random tail vein injection various dose group (four groups) and Lump body injection various dose group (four groups), 5 nude mices of each dosage group, injection dosage respectively for 250mg/kg, 300mg/kg, 350mg/kg and 400mg/kg.Every other day administration, continuous five times.

Naked mouse changes of weight and Survival are observed in experiment, while toxicity of the record with obvious physiological significance, Ru Kang Blighted scurfs, dehydration, drowsiness, incoordination and it is short of breath.If there is Mouse Weight decline>30% Continue more than 3 days or then stop the increase of trial drug dosage because of toxic death, determine that previous group dosage is resistance to for maximum It is within 21 days the experimental observation cycle by dosage, puts to death within the 22nd day animal, autopsy has been visually observed disease-free Become.MTO in tail vein injection is less than 250mg/kg, and the MTO in Lump body injection may be up to 300 mg/kg.Using same dosage, the injection of knurl area looks also more smaller than the toxicity being injected intravenously.

Further investigation revealed that, also there is similar performance in the animal experiment of some other tumours.Embodiment 1 some other quinine class material injections for preparing also have similar performance.Therefore, application selection of the invention Target area administration is main application mode.

3) administration concentrations scope

Experimental animal is lotus HCC nude mice, is divided into negative control group, positive controls and drug research group. The conventional antitumor medicine 5-FU of positive controls injection, the drug research group injection quinine dihydrochloride aqueous solution.Sun Property control group application method be still tail vein injection, negative control group and drug research group are Lump body injection.It is positive Control group application method is still tail vein injection, and negative control group and drug research group are Lump body injection.Each medicine The preparation method of the injection of seminar's (B, C, D, E, F, G group) and the preparation method phase of A2 in table 1 Together, include 2% phenmethylol but contain the quinine dihydrochloride of required various concentrations.Quinine dihydrochloride medication is dense Degree designs anomaly gradient design, and uses logarithm gradient design, i.e., with reference to its IC₅₀Inactive concentration is set The negative control group (A groups) of (5 μM), with its 10²、10³、5×10³、10⁴、8×10⁴、10⁵Experiment is set again Concentration, the injection concentration of quinine dihydrochloride be respectively 0.0005M, 0.005M, 0.025M, 0.05M, 0.4M, 0.5M.Drug research Zu Liu areas injection dosage is less than 300mg quinine dihydrochlorides/kg, and volume injected is by injection Amount and concentration determine that administration number of times is 5 times, and administration frequency is per once two days.Fig. 1 shows two hydrochloric acid Kuis Injection concentration-tumour inhibiting rate the relation curve of peaceful injection.

In Fig. 1, abscissa X is knurl area injection concentration (4 × 10^XμM) index (X), ordinate Y is The tumour inhibiting rate (Y%) measured in thing experiment.In very big concentration range (difference of X is more than 3), i.e., can in general knowledge Prediction more than IC₅₀Tens times, in the range of even thousands of times of Drug level, do not observe and this change in concentration Corresponding pharmacodynamic change, tumour inhibiting rate is less than 25% always.Compared with negative control group, tumour inhibiting rate difference is without statistics Learn meaning (P<0.05).And at 50mM (X=4) place, tumour inhibiting rate mutation rises to 55%, then one (difference of X is equal to 1) is rapidly reached 92% in the range of the Drug level of individual relative narrower, occurs in that breakthrough.In 0.5M Declining occurs again in place, tumour inhibiting rate.

Within whole experimental period, negative control group gross tumor volume shows a rising trend.In the hydrochloric acid of 0.05M-0.5M bis- Compared with negative control group, tumour inhibiting rate difference has statistics to anticipate to drug research group in the range of the injection concentration of quinine Justice (P<0.05).The tumour inhibiting rate of positive control is 75%, and difference has statistics to anticipate compared with negative control group Justice (P<0.05).

Sum it up, when having its Lump body injection concentration only more than a certain critical value, quinine dihydrochloride injection is Show that drug effect, to the extremely sensitive of concentration, then becomes less sensitive again, even tumour inhibiting rate may Occur declining.

General status, changes of weight, the death rate and part that we have also been observed after the injection of animal subject knurl area Denaturation.During testing, each seminar of quinine dihydrochloride injection has no nude mice death, average weight and the moon Property control group changes of weight close to (being dropped by less than 10%), represent that medicine is anti-without obvious toxicity under experimental conditions Should.Compared with positive controls, general status are similar to for seminar.But positive controls average weight declines more (it is more than 20%) greatly, points out a stronger toxic reaction.

During the medication of knurl area, compared with negative control group, in seminar, especially high concentration (>0.4M) Most of injection site has obvious color and luster to change in group, or even with myodegeneration, scope is more in 10mm Within.This points out the partial injury that the application conditions cause myocyte really, injection site to have inflammatory cell to soak Profit, but normal function is not influenceed.After being administered and stopping, the musculature outward appearance of seminar nude mice injection site by Step realizes normalization.The injection site for cuing open the nude mice for killing after stopping to administration for 30 days has carried out check pathological section, Result display seminar is obviously reduced with the difference of negative control group, tends to normal.

Additionally, we also using identical technique study quinine dihydrochloride injection it is different for inoculation time, The thus effect of the otherwise varied knurl body of the development of cancer cell.Drug research group is B groups and C groups.B groups compare C 10 days evenings of group are inoculated with the animal of human liver cancer cell.10 days after last group inoculation, each group selects knurl body respectively Average external volume respectively may be about 28mm³(B groups) and 126mm³8 tumor bearing nude mices injection of (C groups).Lump body injection 0.25M quinine dihydrochlorides, injection dosage is 300mg/kg, and volume injected is determined by injection dosage and concentration. Every other day administration, continuous 6 times.

From experiment the 5th day, each drug research group tumor-bearing mice tumour relative tumour volume was substantially reduced；17 It when tumour relative volume reach minimum；From 32 days, rising occurs in C groups relative tumour volume, and has recurrence Trend；B groups then effect always clearly, temporarily without the trend of recurrence.At 38 days, C groups are than B groups Knurl weight is significantly greater, and the average knurl weight of B groups is close to 0.

This experimental result is illustrated, under the optimum condition in the anti-knurl application that the present invention is provided, two hydrochloric acid Kuis Peaceful injection can be applied to treating malignant tumor, and even can be used for removal completely has the swollen of certain carcinogenic risk Knurl, to prevent it from developing into malignant tumour.

In some documents, quinine dihydrochloride is also considered curing agent.We have selected it has been generally acknowledged that more effective A kind of curing agent (polidocanol) carried out comparative study.At identical conditions, the concentration of polidocanol is From 1.5w/v% until 6w/v% (concentration causes the obvious bubble in knurl area and poisoning symptom occurs), its tumor suppression Rate 30% or so, finds no similar concentration-tumour inhibiting rate relation trend always.The knot of this control experiment Fruit illustrates that composition of the invention is clearly distinguished from the application based on curing agent mechanism.

Similar results are also obtained with other quinine class compound (such as quinine, a quinin hydrochloride) injections.

According to these pharmacodynamic results and local side reaction result, the quinine class compound that the present invention is provided is in tumour Optimum condition in treatment use is as follows：Knurl area is administered, and quinine dihydrochloride injection concentration is 0.05-0.50M, excellent Select 0.15-0.4M, more preferably 0.2-0.3M.

Additionally, another battery of tests shows, pharmaceutical composition of the invention under same concentration, a larger note The amount of penetrating shows more preferable pharmacological effect.It is control dosage if knurl area volume is too big, can be to knurl Area carries out branch's treatment.Each administered volume of injection is necessary for more than the 70% of part volume to be processed, excellent Select more than 110%.When tumour is individual sufficiently small, part volume to be processed is single tumor volume.

Embodiment 3：Application in breast cancer treatment

Experimental animal is lotus breast cancer cell nude mice, is divided into negative control group, positive controls and drug research group. Positive controls tail vein injection routine antitumor medicine adriamycin (7mg/kg), negative control group Lump body injection physiology Salt solution.Research to quinine class compound is carried out first by quinine dihydrochloride.Each drug research group (B, C, D, E, F group) injection preparation method it is identical with the preparation method of A2 in table 1, include 2% phenmethylol, But containing the quinine dihydrochloride of required various concentrations, injection concentration be respectively 0.025M, 0.05M, 0.15M, 0.25M、0.4M.Drug research Zu Liu areas injection dosage is less than 300mg quinine dihydrochlorides/kg, volume injected Determined by injection dosage and concentration, administration number of times is 5 times, administration frequency is per once two days.

Within whole experimental period, negative control group gross tumor volume shows a rising trend；From experiment the 5th day, sun Property control group and each drug research group tumor-bearing mice gross tumor volume are reduced compared with negative control group, and difference has statistics Learn meaning (P<0.05).

Until the 17th day, the Relative tumor proliferation rate of each drug research group tumor-bearing mice from testing the 7th day (T/C) it is respectively less than 40%；From testing the 17th day, B group Relative tumors proliferation rate rises, and has becoming for recurrence Gesture；The Relative tumor proliferation rate (T/C) of C, D, E, F, G group is still reducing, and illustrates that this four groups are being discontinued Still there is obvious tumor growth inhibitory effect afterwards.From testing the 22nd day, C, F group Relative tumor proliferation rate Rise, there is recurrence trend；And D, E group Relative tumor proliferation rate remain close to 0%, effect is very bright always It is aobvious, temporarily without recurrence trend.During testing, suppression ratio D, the E group of positive controls Relative tumor proliferation rate It is late in time, significantly larger in data (40% or so).

The 28th day after administration, the knurl weight of each drug research group tumor-bearing mice is significantly reduced compared with negative control group, And with significant difference (P<0.05), wherein with D, E group tumour inhibiting rate highest (more than 95%)；F groups Tumour inhibiting rate 90.11 ± 2.76%, other group tumour inhibiting rates are less than 85%.Positive controls tumour inhibiting rate is 53%.

Additionally, the present invention provide anti-knurl application in optimum condition under, quinine dihydrochloride injection for The effect for developing otherwise varied knurl body of inoculation time difference thus cancer cell, is also shorter inoculation time Tumour tumor killing effect is better, and it even can be used for having larger carcinogenic risk except can be applied to treatment of cancer The removal completely of nonmalignant tumor, to prevent it to develop into malignant tumor.

During testing, each seminar has no that nude mice is dead, and average weight connects with negative control group changes of weight Closely (it is dropped by less than 10%), represents medicine under experimental conditions without obvious toxic reaction.Seminar is right with the positive Compared according to group, general status are similar to.But positive controls average weight declines bigger (being more than 20%), points out One stronger toxic reaction.

During the medication of target area, compared with negative control group, in seminar, especially most of in F groups There is the change of obvious color and luster injection site, even with myodegeneration, scope is more within 10mm.Enter one The research explanation of step, these denaturation are that quinine dihydrochloride effect is caused.After stopping is administered, experimental group nude mice The musculature outward appearance of injection site is done step-by-step normalization.The note of the nude mice for killing is cutd open within 30 days after stopping to administration Penetrating position has carried out check pathological section, as a result shows that test group is obviously reduced with the difference of negative control group, becomes In normal.

Similar results can also be obtained with other quinine class compound (such as quinine, a quinin hydrochloride) injections. According to these pharmacodynamic results and local side reaction result, the quinine class compound that the present invention is provided is controlled in breast cancer The optimum condition treated in application is as follows：Knurl area is administered, and quinine class compound injection concentration is 0.05-0.50M, excellent Select 0.15-0.4M or 0.2-0.3M.

Additionally, another battery of tests shows that under same concentration, a larger injection volume shows more preferable medicine Effect.It is control dosage if knurl volume is too big, branch's treatment can be carried out.Each administration of injection Volume is necessary for being intended to more than 70%, preferably the 110% of the knurl volume (being tumor volume when compared with tubercle body) for processing More than.

Embodiment 4：Application in liver cancer preventing and treating

Experimental animal is lotus HCC nude mice, and positive controls tail vein injection routine antitumor medicine 5- fluorine is phonetic Pyridine.Research to quinine class compound is carried out first by quinine dihydrochloride.Each drug research group (B, C, D, E, F group) and other experiment conditions it is identical with the method described in embodiment 3.Its experimental result is as follows.

From experiment the 5th day, each drug research group tumor-bearing mice gross tumor volume was reduced compared with negative control group, Statistically significant (the P of difference<0.05).Until the 17th day, each drug research group lotus from testing the 7th day The Relative tumor proliferation rate (T/C) of knurl mouse is respectively less than 40%.From testing the 17th day, higher concentration (0.2M And more than) group Relative tumor proliferation rate (T/C) be respectively less than 15%, illustrate that they still have significantly after drug withdrawal Tumor growth inhibitory effect.

The 28th day after administration, the knurl weight of each drug research group tumor-bearing mice is significantly reduced compared with negative control group, And with significant difference (P<0.05).The tumour inhibiting rate of each drug research group tumor-bearing mice is all higher than>40%, The tumour inhibiting rate of wherein D, E group may be up to more than 95%, and other group tumour inhibiting rates are less than 85%.During testing, Suppression ratio D, E, E, F group of positive controls Relative tumor proliferation rate are late in time, obvious in data Bigger (35% or so), its tumour inhibiting rate is then significantly smaller (72%).

Additionally, the model similar to embodiment 2 is also illustrated, it is excellent in the anti-knurl application that the present invention is provided Under the conditions of choosing, quinine dihydrochloride injection is otherwise varied for the development of inoculation time difference thus cancer cell The effect of knurl body, is also the better shorter tumour tumor killing effect of inoculation time, its except can be applied to oncotherapy, Even can be used for the removal completely of the nonmalignant tumor for having larger carcinogenic risk, to prevent it to develop into malignant tumor.

During being tested more than, each seminar has no animal dead, and average weight becomes with negative control group body weight Change close to (being dropped by less than 10%), represent medicine under experimental conditions without obvious toxic reaction.Seminar and sun Property control group compare, general status be similar to.But it is bigger that transplantable tumor tests positives control group average weight decline (being more than 20%), point out a stronger toxic reaction.

Similar results are also obtained with other quinine class compound (such as quinine, a quinin hydrochloride) injections.According to These pharmacodynamic results and local side reaction result, the quinine class compound that the present invention is provided is in liver cancer treatment application In optimum condition it is as follows：Knurl area is administered, and quinine class compound injection concentration is 0.05-0.50M, preferably 0.15-0.4M or 0.2-0.3M.

Additionally, another battery of tests shows that under same concentration, a larger injection volume shows more preferable medicine Effect.It is control dosage if knurl volume is too big, branch's treatment can be carried out.Each administration of injection Volume is necessary for more than 70%, preferably the 110% of part knurl volume to be processed (being tumor volume when compared with tubercle body) More than.

Embodiment 5：Application in pancreas cancer controlling

Experimental animal is pancreatic tumor borne cell nude mice.Positive controls tail vein injection routine antitumor medicine Ji Xita Shore (100mg/kg).Research to quinine class compound is carried out first by quinine dihydrochloride.Each drug research The method of breast cancer treatment experiment in group (B, C, D, E, F group) and other experiment conditions and embodiment 3 It is identical.Its experimental result is as follows.

From experiment the 5th day, each drug research group tumor-bearing mice gross tumor volume was reduced compared with negative control group, Statistically significant (the P of difference<0.05).Until the 17th day, each drug research group lotus from testing the 7th day The Relative tumor proliferation rate (T/C) of knurl mouse is respectively less than 40%.From testing the 17th day, C, D, E group Relative tumor proliferation rate (T/C) is respectively less than 15%, illustrates that they still have obvious tumour growth to press down after drug withdrawal Effect processed.

The 28th day after administration, the knurl weight of each drug research group tumor-bearing mice is significantly reduced compared with negative control group, And with significant difference (P<0.05).The tumour inhibiting rate of each drug research group tumor-bearing mice is all higher than>40%, The tumour inhibiting rate of wherein D, E group may be up to more than 90%.Other group tumour inhibiting rates are less than 85%.During testing, Suppression ratio D, E, E, F group of positive controls Relative tumor proliferation rate are late in time, obvious in data Bigger (35% or so), its tumour inhibiting rate is then significantly smaller (62%).

Additionally, the model similar to embodiment 2 is also illustrated, it is excellent in the anti-knurl application that the present invention is provided Under the conditions of choosing, quinine dihydrochloride injection is otherwise varied for the development of inoculation time difference thus cancer cell The effect of knurl body, is also the better shorter tumour tumor killing effect of inoculation time, its except can be applied to treatment of cancer, Even can be used for the removal completely of the nonmalignant tumor for having larger carcinogenic risk, to prevent it to develop into malignant tumor.

Some other the quinine class compounds (such as quinine, a quinin hydrochloride) prepared using embodiment 1 are injected Agent, it is also possible to obtain similar results.According to these pharmacodynamic results and local side reaction result, the present invention is provided Optimum condition of the quinine class compound in treatment of pancreatic cancer application it is as follows：Knurl area is administered；Quinine class compound Injection concentration is 0.05-0.50M, preferably 0.15-0.4M or 0.2-0.3M.

Embodiment 6：Application in thyroid cancer preventing and treating

Experimental animal is lotus thyroid carcinoma cell nude mice, positive controls tail vein injection routine antitumor medicine Ah mould Plain (4mg/kg), negative control group Lump body injection physiological saline.To the research of quinine class compound first by one Quinin hydrochloride is carried out.The preparation method and table 1 of the injection of each drug research group (B, C, D, E, F group) The preparation method of middle B2 is identical, includes 2% phenmethylol, but the quinin hydrochloride containing required various concentrations, Injection concentration is respectively 0.025M, 0.05M, 0.15M, 0.25M, 0.4M.Other experiment conditions and reality Breast cancer treatment experiment is identical in applying example 3.Its experimental result is as follows.

In administration phase, the Relative tumor proliferation rate of each drug research group tumor-bearing mice declines；From testing the 17th day, Each drug research group Relative tumor proliferation rate beyond B groups very low level (<35%).Test the 22nd day Rise, close to 20%, effect clearly, temporarily becomes C, D, E group Relative tumor proliferation rate without recurrence always Gesture.During testing, each drug research beyond the suppression ratio B groups of positive controls Relative tumor proliferation rate Group is late in time, significantly larger in data (40% or so).

The 28th day after administration, the knurl weight and negative control group phase of each drug research group tumor-bearing mice beyond B groups Than significantly reducing, and with significant difference (P<0.05), wherein with D group tumour inhibiting rates highest (more than 85%), E groups tumour inhibiting rate is less than 80%.Positive controls tumour inhibiting rate is 61%.

In addition, the work of the injection knurl body otherwise varied for the development of inoculation time difference thus cancer cell With, be also the better shorter tumour tumor killing effect of inoculation time, its except can be applied to treatment of cancer, even Can be used for the removal completely of the non-malignant tumors for having larger carcinogenic risk, to prevent it to develop into malignant tumor.

Some other quinine class compound (such as quinine, quinine dihydrochloride) injections prepared using embodiment 1, Similar results can also be obtained.According to these pharmacodynamic results and local side reaction result, the Kui that the present invention is provided Peaceful optimum condition of the class compound in treatment of thyroid carcinoma application is as follows：Knurl area is administered；Quinine class compound is noted Concentration is penetrated for 0.05-0.50M, preferably 0.15-0.4M or 0.2-0.3M.

In addition, another battery of tests shows that under same concentration, a larger injection volume shows more preferable drug effect.Such as Fruit knurl volume is too big, is control dosage, can carry out branch's treatment.Each administered volume of injection must It must be more than 70%, preferably more than the 110% of part knurl volume to be processed (being tumor volume when compared with tubercle body).

Embodiment 7：Application in prostate cancer preventing and treating

Experimental animal is lotus prostate gland cancer cell nude mice, positive controls tail vein injection routine antitumor medicine Ah mould Plain (4mg/kg), negative control group Lump body injection physiological saline.To the research of quinine class compound first by one Quinin hydrochloride is carried out.Each drug research group (B, C, D, E, F group) and treatment of thyroid carcinoma in embodiment 6 Experiment is identical, and other experiment conditions are identical with breast cancer treatment experiment in embodiment 3.Its experimental result is as follows. Within whole experimental period, negative control group gross tumor volume shows a rising trend；From experiment the 5th day, positive control Each drug research group tumor-bearing mice gross tumor volume beyond group and B groups is reduced compared with negative control group, difference Statistically significant (P<0.05).

In administration phase, the Relative tumor proliferation rate of each drug research group tumor-bearing mice declines；From testing the 17th day, Each drug research group Relative tumor proliferation rate beyond B groups very low level (<40%).Test the 22nd day Rise, C, D group Relative tumor proliferation rate are less than 15%, effect always clearly, temporarily without the trend of recurrence. During experiment, each drug research group beyond the suppression ratio B groups of positive controls Relative tumor proliferation rate when Between upper evening, significantly larger in data (being more than 35%).

The 28th day after administration, the knurl weight and negative control group phase of each drug research group tumor-bearing mice beyond B groups Than significantly reducing, and with significant difference (P<0.05), wherein with C, D group tumour inhibiting rate highest (90% with On), other group tumour inhibiting rates are less than 80%.Positive controls tumour inhibiting rate is 63%.

Additionally, the injection knurl body otherwise varied for the development of inoculation time difference thus cancer cell Effect, be also the better shorter tumour tumor killing effect of inoculation time, except can be applied to relatively late prostate cancer and Early prostate cancer, even can be used for the removal completely of the nonmalignant tumor for having larger carcinogenic risk, to prevent it Develop into malignant tumor.

Some other the quinine class compounds (such as quinine, quinine dihydrochloride) prepared using embodiment 1 are injected Agent, it is also possible to obtain similar results.According to these pharmacodynamic results and local side reaction result, the present invention is provided Optimum condition of the quinine class compound in prostate cancer therapy application it is as follows：Knurl area is administered；Quinine class chemical combination Thing injection concentration is 0.05-0.50M, preferably 0.15-0.4M or 0.2-0.3M.

Embodiment 8：Application in treatment of nasopharyngeal carcinoma

Experimental animal is lotus nasopharyngeal carcinoma cell nude mice, positive controls tail vein injection routine antitumor medicine 5- fluorine Pyrimidine.Research to quinine class compound is carried out first by quinine dihydrochloride.Each drug research group (B, C, D, E, F group) and other experiment conditions with embodiment 3 breast cancer treatment test method it is identical.In fact Test result as follows.

The 28th day after administration, the knurl weight of each drug research group tumor-bearing mice is significantly reduced compared with negative control group, And with significant difference (P<0.05).The Relative tumor tumour inhibiting rate of each drug research group tumor-bearing mice is all higher than >40%, the tumour inhibiting rate of wherein higher concentration (0.2M and more than) group may be up to more than 95%, other group tumour inhibiting rates Less than 85%.During testing, suppression ratio D, E of positive controls Relative tumor proliferation rate, E, F group exist Evening, significantly larger in data (30% or so) on time；Tumour inhibiting rate average value is 68%.

Some other quinine class compound (such as quinine, a quinin hydrochloride) injections prepared using embodiment 1, Similar results can also be obtained.According to these pharmacodynamic results and local side reaction result, the Kui that the present invention is provided Peaceful optimum condition of the class compound in treatment of nasopharyngeal carcinoma application is as follows：Knurl area is administered；Quinine class compound injection Concentration is 0.05-0.50M, preferably 0.15-0.4M or 0.2-0.3M.

Embodiment 9：Application in lung cancer therapy

Experimental animal is lotus lung carcinoma cell nude mice, and positive controls tail vein injection routine antitumor medicine 5- fluorine is phonetic Pyridine.Research to quinine class compound is carried out first by quinine dihydrochloride.Each drug research group (B, C, D, E, F group) and other experiment conditions with embodiment 3 breast cancer treatment test method it is identical.Its experiment knot Fruit is as follows.

From experiment the 5th day, each drug research group tumor-bearing mice gross tumor volume was reduced compared with negative control group, Statistically significant (the P of difference<0.05).Until the 17th day, each drug research group lotus from testing the 7th day The Relative tumor proliferation rate of knurl mouse is obviously reduced.From testing the 17th day, higher concentration (0.2M and more than) group Relative tumor proliferation rate (T/C) be respectively less than 15%, illustrate that they still have obvious tumour growth after drug withdrawal Inhibition.

The 28th day after administration, the knurl weight of each drug research group tumor-bearing mice is significantly reduced compared with negative control group, And with significant difference (P<0.05).The Relative tumor tumour inhibiting rate of each drug research group tumor-bearing mice is all higher than >40%, the tumour inhibiting rate of wherein higher concentration (0.2M and more than) group may be up to more than 95%, other group tumour inhibiting rates Less than 85%.During testing, suppression ratio D, E of positive controls Relative tumor proliferation rate, E, F group exist Evening, significantly larger in data (30% or so) on time；Positive controls tumour inhibiting rate is 54%.

Some other the quinine class compounds (such as quinine, a quinin hydrochloride) prepared using embodiment 1 are injected Agent, it is also possible to obtain similar results.According to these pharmacodynamic results and local side reaction result, the present invention is provided Optimum condition of the quinine class compound in lung cancer therapy application it is as follows：Knurl area is administered；Quinine class compound is noted Concentration is penetrated for 0.05-0.50M, preferably 0.15-0.4M or 0.2-0.3M.

Embodiment 10：Application in treatment of colon cancer

Experimental animal is lotus colon cancer cell nude mice.Positive controls tail vein injection routine antitumor medicine cis-platinum. Research to quinine class compound is carried out first by quinine dihydrochloride.Each drug research group (B, C, D, E, F groups) and other experiment conditions with embodiment 3 breast cancer treatment test method it is identical.Its experimental result It is as follows.

The 28th day after administration, the knurl weight of each drug research group tumor-bearing mice is significantly reduced compared with negative control group, And with significant difference (P<0.05).The Relative tumor tumour inhibiting rate of each drug research group tumor-bearing mice is all higher than >40%, the tumour inhibiting rate of wherein higher concentration (0.2M and more than) group may be up to more than 95%, other group tumour inhibiting rates Less than 85%.During testing, suppression ratio D, E of positive controls Relative tumor proliferation rate, E, F group exist Evening, significantly larger in data (30% or so) on time；Tumour inhibiting rate average value is 75%.

Some other the quinine class compounds (such as quinine, a quinin hydrochloride) prepared using embodiment 1 are injected Agent, it is also possible to obtain similar results.According to these pharmacodynamic results and local side reaction result, the present invention is provided Optimum condition of the quinine class compound in treatment of colon cancer application it is as follows：Knurl area is administered；Quinine class compound Injection concentration is 0.05-0.50M, preferably 0.15-0.4M or 0.2-0.3M.

Claims

1. a kind of pharmaceutical composition for treating tumour, it includes quinine class compound and can pharmaceutically connect The excipient or carrier received, and be the formulation for being suitable for being administered through knurl area, wherein the medicine when being administered through knurl area The concentration of quinine class compound is 0.05-0.5M in compositions.

2. the pharmaceutical composition described in claim 1, wherein the concentration of the quinine class compound is 0.15-0.4M, preferably 0.2-0.3M.

3. the pharmaceutical composition described in claim 1, wherein when knurl area is administered, the volume of the injection Not less than the tumor mass volume for being intended to process 70%, preferably not less than 110%.

4. the pharmaceutical composition described in claim 1, wherein the quinine class compound includes one below kind Or it is various：Quinine, quinine isomers and their derivative.

5. the pharmaceutical composition described in claim 4, wherein the derivative is acid-addition salts, including it is inorganic Acid-addition salts and organic acid addition salt, the inorganic acid addition salt preferably include one below kind or various：Hydrochloric acid Salt, hydrobromate, hydriodate, dihydrochloride, dihydrobromide, two hydriodates, disulfate, sulphur Hydrochlorate, phosphate, and the organic acid addition salt preferably includes one or more of：Bicarbonate or carbonic acid Salt, ethyl carbonate salt, formates, acetate, Chinese holly edge hydrochlorate, tannate.

6. the pharmaceutical composition described in claim 1, wherein the pharmaceutically acceptable excipient or load Body includes water, the miscible organic solvent of water or their mixture, and the miscible organic solvent of water is preferred Including one below kind or various：Ethanol, propane diols, PEG, isopropanol.

7. the pharmaceutical composition described in claim 1, its also comprising anodyne, other antineoplastics and/ Or synergist, the anodyne preferably includes one below kind or various：Phenmethylol, procaine hydrochloride, three The chlorine tert-butyl alcohol, hydrochloric acid benefit card, and the synergist preferably includes curing agent and/or immunologic adjuvant.

8. the pharmaceutical composition described in claim 7, wherein the curing agent includes one below kind or various： Lauromacrogol, sodium morrhuate, EO, polidocanol, bleomycin A5.

9. the pharmaceutical composition described in claim 1, wherein the tumour includes non-malignant tumors and pernicious swollen Knurl, the malignant tumour includes breast cancer, liver cancer, cancer of pancreas, thyroid cancer, prostate cancer, lung cancer, head Neck cancer, colon cancer, nasopharyngeal carcinoma.