CN106860459A - Application, method and pharmaceutical composition of the quinolines material in treatment thyroid gland and mammary gland disease - Google Patents

Application, method and pharmaceutical composition of the quinolines material in treatment thyroid gland and mammary gland disease Download PDF

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Publication number
CN106860459A
CN106860459A CN201510923813.2A CN201510923813A CN106860459A CN 106860459 A CN106860459 A CN 106860459A CN 201510923813 A CN201510923813 A CN 201510923813A CN 106860459 A CN106860459 A CN 106860459A
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quinolines
injection
pharmaceutical composition
group
concentration
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邹方霖
卢星
邹礼常
王建霞
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Chengdu Kuachang Science and Technology Co Ltd
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Chengdu Kuachang Science and Technology Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/49—Cinchonan derivatives, e.g. quinine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4706—4-Aminoquinolines; 8-Aminoquinolines, e.g. chloroquine, primaquine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4709—Non-condensed quinolines and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/517—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with carbocyclic ring systems, e.g. quinazoline, perimidine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca

Abstract

The application in treatment thyroid gland, mammary gland disease and method the present invention relates to quinolines material, and the pharmaceutical composition comprising quinolines material.

Description

Application, method and pharmaceutical composition of the quinolines material in treatment thyroid gland and mammary gland disease
Technical field
The application in treatment thyroid gland, mammary gland disease and method the present invention relates to quinolines material, and bag Pharmaceutical composition containing quinolines material.
Background technology
Many quinolines materials are antiparasitic, especially anti-malarial agents.Quinolines material is outside antimalarial Using much being carried out with composition forms.This based composition is once applied to treat various diseases, for example Hemorrhoid, rhinitis, hemangioma, facial paralysis etc..It is generally believed that other components (such as urethane) in composition Not only some drug effects also can in the treatment be played with hydrotropy.Thus, include quinolines thing for specified disease The pharmaceutical composition of matter is always among research.For example, Chinese patent 87104056 discloses a kind of hydrochloric Six components compositions " injection for whole prolapse haemorrhoids " of quinine and quinine dihydrochloride, for removing hemorrhoid and being closed with hemorrhoid The anal fissure of connection, rectal polyp and papilla fiber histioma.But in the composition, quinolines material concentration More than 0.47M, and contain urethane.Chinese patent 200910103187.7 is combined there is provided a kind of nine component Thing is used to treat hemorrhoid and tumour.The concentration very little (being less than 0.05M) 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 application.It is Chinese special Profit 96110932 discloses a kind of composition comprising a quinin hydrochloride and Anodynine, and it is used for Treatment of Hyperthyroidism. However, Anodynine side effect is very strong, in developed country, oneself is eliminated.
Possible application of the quinolines material in terms of anti-knurl once received very many concerns.Prior art is provided The anti-tumor activity condition of some quinolines, such as Chinese patent of following application number：87106996、 200710119982.6th, 201210111920.1 etc., and following summary：Wang Jing etc., the anti-of quinoline swells Tumor activity progress, World Notes on Antibiotics, 2013, Vol.34, No.2, pp60-70.In fact, In the in vitro test with tumour cell as model, some quinolines materials 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 quinolines material (for example Quinine dihydrochloride) injection concentration improve about 500 times (0.025M), effective anti-knurl effect is also not observed (tumour inhibiting rate is less than 40%), does not show the drug effect more considerably higher than existing antitumor medicine.In the prior art, its The tumour inhibiting rate of its quinine class medicine is not also high.In the last few years, people just no longer regarded them as a kind of competitive Antitumor medicine, and be treated as anti-knurl activity increase agent and studied.For example, No. 201310459568.5 China Patent application is exactly resistance to the multiple medicine that antitumor medicine (vincristine class medicine) is used to reduce cell by quinine The property of medicine is so that antitumor medicine reaches preferable drug effect.
Current thyroid gland, the treatment of mammary gland disease, the mainly change of operation, radiation cure and other medicines Treat.Operation is damaged to body, and the mammary gland attractive in appearance on some influences and thyroid injury are frustrating, It is set to become a kind of painful choice.The side effect of radiation cure is also clearly.Existing chemotherapy is substantially Systemic administration, there is systemic side effects risk very high.For example, existing antitumor medicine is in interior killing cancer on a large scale While cell, a large amount of normal cells are also killed, immune system is seriously damaged, and curative effect is not yet It is satisfactory.
The content of the invention
Goal of the invention of the invention is to provide one kind, and more effectively thyroid gland, breast are treated in smelting compared with prior art The pharmaceutical composition and treatment method of gland disease.
According to an aspect of the present invention, it provides a kind of medicine group for treating thyroid gland, mammary gland disease Compound, it includes quinolines material and pharmaceutically acceptable excipient, and to be suitable for through thyroid gland, breast Gland disease target area administration formulation, wherein when being administered through target area in described pharmaceutical composition quinolines material it is dense It is 0.05-0.5M to spend, preferably 0.1-0.4M, more preferably 0.1-0.3M.
According to another aspect of the present invention, it provides quinolines material in treatment thyroid gland, mammary gland disease Application, wherein the quinolines material is included in pharmaceutical composition, described pharmaceutical composition includes materia medica Acceptable excipient and be the formulation for being suitable for being administered through target area, wherein when target area is administered, the medicine group The concentration of quinolines material is 0.05-0.5M, more preferably preferably 0.1-0.4M, 0.1-0.3M in compound.
According to a further aspect of the invention, it provides a kind of method for treating thyroid gland, mammary gland disease, its Including comprising the quinolines material and pharmaceutically medicine of acceptable excipient is administered to the target area for having this to need Composition, wherein described pharmaceutical composition are the formulation for being suitable for being administered through target area, and when the target area is administered, In described pharmaceutical composition the concentration of quinolines material be 0.05-0.5M, preferably 0.1-0.4M, more preferably 0.1-0.3M。
Within the scope of the invention, when target area is administered, the administered volume of described pharmaceutical composition is not less than desire 70%, preferably not less than the 110% of the 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, in addition to quinolines material and pharmaceutically acceptable excipient or carrier, The composition can also further include anodyne, other antineoplastics and/or synergist.
Within the scope of the invention, the thyroid gland, mammary gland disease include breast cancer, mammary gland non-malignant tumors, The non-struma of mammary gland is big or non-knurl hyperplasia, galactoncus, thyroid cancer, thyroid gland non-malignant tumors, thyroid gland are non- Struma is big or non-knurl hyperplasia, thyroid cyst, hyperthyroidism 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 dynamic smaller)；Compared with existing systemic administration is treated, can be under the toxic and side effect for minimizing, conspicuousness ground Improve curative effect；Compared with existing local application, especially curing agent (such as absolute ethyl alcohol) are treated, with disease The compatibility become 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 × 10XμM) index (X), and ordinate Y is the tumour inhibiting rate (Y%) measured in zoopery.
Fig. 2 is the injection concentration-tumour inhibiting rate relation curve of primaquine injection, and wherein abscissa X is noted for knurl area Penetrate concentration (4 × 10XμM) index (X), and ordinate Y is the tumour inhibiting rate (Y%) measured in zoopery.
Specific embodiment
Within the scope of this invention, term " quinolines material " refers to one or more quinoline containing quinoline ring Derivative, it is preferably selected from in the following group：Chloroquine (chloroquine), HCQ quinoline, ammonia first quinoline, ammonia phenol Quinoline, quinine, nitroquine, PIPERAQUINE, primaquine (primaquine), pamaguine, pentane quinoline and its isomers Pharmaceutically acceptable salt.
For quinine, its isomers for example can be quinindium, cinchonine and cinchonidine.
The pharmaceutically acceptable salt of quinolines material includes acid-addition salts, such as inorganic acid addition salt and organic Acid-addition salts.The example of inorganic acid addition salt is included but is not limited to：Halogen acid salt (such as hydrochloride (such as hydrochloric acid Kui Rather), hydrobromate, hydriodate, dihydrochloride (such as quinine dihydrochloride), dihydrobromide, two hydroiodic acids Salt), disulfate, sulfate, phosphate (such as chloroquine diphosphate, piperaquine phosphate, primaquine phosphate), etc.. The example of organic acid addition salt is included but is not limited to：Bicarbonate or carbonate, ethyl carbonate salt, formates, Acetate, Chinese holly edge hydrochlorate or tannate.
Above-mentioned quinolines material can be used alone, and also can be combined and uses, and such as quinin hydrochloride and other quinoline spread out Biological combination, the composition of quinine dihydrochloride and other quinolines, primaquine and other quinolines The composition of composition, chloroquine and other quinolines, etc..
Within the scope of this invention, pharmaceutically acceptable excipient (or solvent) can be water, water Miscible organic solvent or the vehicle system comprising water and the miscible organic solvent of water.Water is miscible Organic solvent includes one below kind or various：Ethanol, propane diols, polyethylene glycol (PEG), isopropanol. Wherein PEG can optionally employ PEG, such as PEG200, PEG300 and PEG400 of the molecular weight less than 600 Deng.The composition is free of urethane and antipyrine.
Within the scope of the invention, in addition to quinolines material and pharmaceutically acceptable excipient, the combination Thing can also further include anodyne.The anodyne can be well known by persons skilled in the art any appropriate Person, to mitigate the pain of patient, such as phenmethylol, procaine hydrochloride, anesin, hydrochloric acid benefit Card etc..
Within the scope of the invention, in addition to quinolines material and pharmaceutically acceptable excipient, the combination Thing can also further include other antineoplastics.The antineoplastic can be those skilled in the art Any appropriate person for knowing, to further enhance antitumor activity, such as including adriamycin, 5-FU, Ji Xi His shore, cis-platinum, qinghaosu and its derivative etc..
Within the scope of the invention, in addition to quinolines material and pharmaceutically acceptable excipient, the combination Thing can also further include synergist.The synergist can be well known by persons skilled in the art any appropriate Person, to further enhance the effect for the treatment of thyroid gland, mammary gland disease, 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 ill area The necrosis of interior enlargement, reduce and fibrosis, it for example can be Lauromacrogol, sodium morrhuate, monoethanolamine oleic acid Ester, polidocanol 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 " target area administration " refers to that medicine is added directly into the local interval of drug target Relatively centralized It is interior.For example, when disease is enlargement, target area is the enlargement area including lump and block week.When lump is tumour When, target area is including knurl body and Liu Zhouliu areas；When lump is non-struma block (or accretion prism), target area As include the enlargement area in lump (or accretion prism) and block (or accretion prism) week；When lump is tumour, target Area is capsule.When disease is hyperthyroidism, target area is thyroid gland.When target area is administered, medicine of the invention Compositions are the formulation for being suitable for target area administration, for example, can be injection etc., and quinolines material is in the target Area administration pharmaceutical composition in concentration be 0.05-0.5M (Mol/L), preferably 0.1-0.4M, more preferably It is 0.1-0.3M.In addition, when target area is administered, the administered volume of described pharmaceutical composition is not less than being intended to process it 70%, preferably not less than the 110% of volume, so that the pharmaceutical composition comprising quinolines material can infiltrate Target spot and its peripheral region, realize therapeutic action higher.When disease is enlargement, if its lump is (for example Tumour, non-struma block, capsule) volume it is sufficiently small, then the volume is the above-mentioned volume for being intended to process.For The larger enlargement of lump and hyperthyroidism, then need to be allocated as the medication in batches of multiple parts, then the part body Product 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 quinolines material for required concentration in water-soluble concentration range, such as concentration is less than The quinine dihydrochloride of 0.35M, and the miscible strong anodyne of water, other antineoplastics being optionally present And/or synergist, obtain corresponding injection by they are directly added into water for injection.
Method two：Solid phase quinolines material for required concentration outside water-soluble concentration range, such as concentration is big In a quinin hydrochloride of 0.15M, and the miscible difference of water being optionally present anodyne, other are antitumor Medicine and/or synergist, then first prepare required vehicle system (such as 9% ethanol solution), then will be above-mentioned Solid matter is added in the vehicle system, you can obtain the injection of the quinolines material of concentration comprising needed for.
Method three：When the anodyne for being insoluble in water, other antineoplastics and/or synergist need to be added, then Can for example prepare by the following method：
A. the quinolines material aqueous solution, or the solution for preparing required vehicle system and quinolines material are prepared；
B. vehicle system needed for preparing and anodyne, other antineoplastics and/or the synergist for being insoluble in water Solution；
C. the solution that a and b is prepared is mixed.
By above-mentioned these methods, the various injections comprising quinolines material can be prepared.For example, different injections Change in agent includes：Containing different quinolines materials, the quinolines material containing various concentrations, containing different water Miscible organic solvent, the miscible organic solvent of water containing various concentrations, containing variety classes and concentration only Pain agent, other antineoplastics and/or synergist, etc..But change anyway, quinolines material is in knurl Total concentration in the pharmaceutical composition of area's administration should remain 0.05-0.5M (Mol/L), preferably 0.1-0.4M, More preferably 0.1-0.3M.
If it does, the concentration of anodyne is 0-3% (weight)；The concentration of other antineoplastics is 0-0.15M； The concentration of synergist is 0-3% (weight)
Certainly, the injection concentration comprising quinolines material (such as quinine dihydrochloride) needed for the present invention can also lead to Diluting high-concentration quinolines material parenteral solution (such as containing the parenteral solution of 25W/V% quinine dihydrochlorides) is crossed to obtain.
Within the scope of the invention, pharmaceutical composition comprising quinolines material can also be before knurl area is administered 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 quinolines material at administration, improve its life Thing availability, its can for example mix micro particles as slow-released carrier or matrix, nano-particle, micella and 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 breast cancer cell And human thyroid cancer cell (SW579) (MDA-MB231).Research method and knot in embodiments of the invention Fruit is also applied for quinolines material other tumour cells of anti-tumor activity to it.Treat that tumour is long to 50-100mm3 When, animal subject is carried out at random using the softwares of PEMS 3.2 (establishment of West China HSPH of Sichuan University) District's groups, respectively 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=Vt/V0, V in formula0It is the packet administration same day (i.e. the One day) measurement obtained by gross tumor volume, VtGross tumor volume during to measure each time.
Relative tumor proliferation rate computing formula is as follows：T/C (%)=TRTV/CRTV× 100, in formula：TRTVIt is the positive The RTV of control group or drug research group, CRTVIt 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%, in formula： 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 quinolines material injection
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 preparation A1, A5, A6, B1, B4, C1, D1 and F1 are according to similar to the above Method is prepared.
The injection concentration of the required quinolines material (such as quinine dihydrochloride) in the present invention, also can be by dilution High concentration quinolines material (such as quinine dihydrochloride of W/V percentages 25%) parenteral solution is obtained.
Embodiment 2：The optimum condition research of application
The optimum condition of application of the quinolines material of the invention in thyroid gland, medicinal breast disease treatment, exists first Studied in anti-knurl experiment (external inhibiting tumor cell experiment and transplantable tumor animal model experiment).
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 living Property, refer to that there is certain anti-homophylic property of knurl compared with existing clinical medicine.A kind of material is such as real in vitro There is relatively low 503nhibiting concentration (to be abbreviated as IC in testing50, 50 μM are less than in generally suppressing reaction at 72 hours), Or inhibition rate of tumor growth higher (abbreviation tumour inhibiting rate, typically larger than 40% but small are shown in zoopery In 75%), just it is considered as having similarity with existing clinical medicine, so as to show anti-tumor activity, the thing Matter is also just referred to as active material.Term " reactive conditions " refers to that the something that makes that researcher provides shows it The condition of anti-tumor activity.For example, making the tumour inhibiting rate of quinolines material 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.
1) anti-tumor activities research
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 Study carefully group group and be respectively the quinolines substance solution of the various concentrations diluted with higher concentration mother liquor (in a μM ol/l In level).These mother liquors be respectively embodiment 1 preparation containing the quinin hydrochlorides of 0.25M mono-, the hydrochloric acid of 0.25M bis- The injection of quinine, 0.15M quinines, 0.15M chloroquine diphosphates and 0.1M primaquine.
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 .37 DEG C of the serum free medium containing 0.2mg/ml MTT of 200 μ l Fresh is added to continue to cultivate 4h. It is careful to abandon supernatant, and 200 μ l DMSO are added, after being mixed with miniature ultrasonic oscillator, with examination on ELIASA Wavelength is tested for 570nm, reference wavelength is that 450nm determines OD value.It is calculated as follows drug on tumor thin The inhibiting rate of intracellular growth：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 quinolines material concentration in composition.Generally speaking, they all μM concentration level, system Meter does not see significant difference on learning.And the IC of positive control50Then between 0.01-0.5 μM.
Table 2
*：The total concentration of quinolines material.
Further test and also demonstrate, the organic solvent in concentration range, the analgesic introduced in the present composition Agent, synergist have no negative effect to the anti-tumor activity of quinolines material.
2) application method research
Experimental animal is lotus breast cancer cell nude mice, is divided into negative control group, positive controls and drug research group. Negative control group injecting normal saline, conventional antitumor medicine adriamycin (4mg/kg) of positive controls injection, medicine Quinine dihydrochloride solution is injected by seminar.When tail vein injection is small, in, heavy dose of concentration be 0.025M's During the quinine dihydrochloride aqueous solution, although we have also made some technical improvement, eligible result still with existing skill The result of art is consistent, and tumour inhibiting rate is less than 30%.And reach 50% as the tumour inhibiting rate of the adriamycin 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 two hydrochloric acid The animal tail vein injection of quinine and Lump body injection maximum tolerated dose (Maximum telareted dose, MTO), method is as follows：
Experimental animal is lotus breast cancer cell nude mice, and tail vein injection various dose group (four groups) is grouped at random With Lump body injection various dose group (four groups), 5 nude mices of each dosage group, injection dosage is respectively 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 300mg/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.Implement Some other quinolines material injections prepared by example 1 also have similar performance.Therefore, application choosing of the invention It is main application mode to have selected target area administration.
3) administration concentrations scope
Experimental animal is lotus breast cancer cell nude mice, is divided into negative control group, positive controls and drug research Group.Negative control group injecting normal saline, conventional antitumor medicine adriamycin (4mg/kg) of positive controls injection, Drug research group injects the quinine dihydrochloride aqueous solution of various concentrations.Positive controls application method is still tail vein Injection, negative control group and drug research group are Lump body injection.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 Need various concentrations quinine dihydrochloride, injection concentration be respectively 0.025M, 0.05M, 0.15M, 0.25M, 0.4M、0.5M.Drug research Zu Liu areas injection dosage be less than 300mg quinine dihydrochlorides/kg, volume injected by Injection dosage and concentration determine that administration number of times is 5 times, and administration frequency is per once two days.Fig. 1 shows two The injection concentration of quinin hydrochloride injection-tumour inhibiting rate relation curve.
In Fig. 1, abscissa X is knurl area injection concentration (4 × 10XμM) 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 45%, then one (difference of X is equal to 1) is rapidly reached more than 85% in the range of the Drug level of individual relative narrower, occurs in that breakthrough. At 0.4M, declining occurs again in tumour inhibiting rate.
Within whole experimental period, negative control group gross tumor volume shows a rising trend.In the hydrochloric acid of 0.05M-0.4M 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 53%, and difference has statistics to anticipate compared with negative control group Justice (P<0.05).
As shown in Fig. 2 carrying out above-mentioned experiment with primaquine injection, similar result is obtained.
Sum it up, when having its Lump body injection concentration only more than a certain critical value, quinine dihydrochloride or primaquine Injection just shows that drug effect, to the extremely sensitive of concentration, then becomes less sensitive again, even presses down Ratio of outflow is likely to occur decline.
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) In group most of injection site have obvious color and luster change, 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 12 days evenings of group are inoculated with the animal of human breast cancer cell.10 days after last group inoculation, each group selects knurl respectively Body average external volume respectively may be about 25mm3(B groups) and 103mm38 tumor bearing nude mices injection of (C groups).Knurl body is noted 0.25M quinine dihydrochlorides are penetrated, injection dosage is 300mg/kg, and volume injected is true by injection dosage and concentration It is fixed.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.
The specification of a model that this experiment is provided, under the optimum condition in the anti-knurl application that the present invention is provided, two Quinin hydrochloride injection can be applied to treating malignant tumor, and even can be used for removal completely has certain carcinogenic risk Tumour, 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.
Also obtained with some other the quinolines material injections (such as the injection in table 1) prepared in embodiment 1 Obtain similar results.Additionally, in the examples below, when quinolines material is applied to other thyroid glands, mammary gland disease Similar results are also obtained during the zoopery of disease.
According to these pharmacodynamic results and local side reaction result, the quinolines material that the present invention is provided is in first shape Optimum condition in gland, medicinal breast disease treatment application is as follows：Target area is administered, and quinolines material injection concentration is 0.05-0.50M, preferably 0.1-0.4M, more preferably 0.1-0.3M.
Additionally, another battery of tests shows, quinolines material injection of the invention under same concentration, one Larger injection volume shows more preferable pharmacological effect.It is control dosage if Gross Target Volume is too big, Branch's treatment can be carried out to target area.Each administered volume of injection is necessary for the 70% of part volume to be processed Above, preferably more than 110%.When the target block of target area is lump (such as tumour, non-struma block) and its individual foot Enough hours, part volume to be processed is single lump volume.
Embodiment 3：Application in medicinal breast disease treatment
Clinically, most common mammary gland disease is mammary gland enlargement, including tumour and non-struma it is big.This implementation In example, the application study to quinolines material 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 required various concentrations quinine dihydrochloride, its injection concentration be respectively 0.025M, 0.05M, 0.15M, 0.25M、0.4M.Injection dosage is less than 300mg quinine dihydrochlorides/kg animals, and volume injected is by injection dosage Determine with concentration.
1. tumor of breast 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 Physiological saline.Drug research Zu Liu areas injection dosage is less than 300mg quinine dihydrochlorides/kg, and volume injected is by noting Penetrate dosage and concentration determines, 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 the trend of recurrence； The Relative tumor proliferation rate (T/C) of C, D, E, F group is still reducing, and illustrates that this four groups still have after drug withdrawal There is obvious tumor growth inhibitory effect.From testing the 22nd day, C, F group Relative tumor proliferation rate rise, and have Recurrence trend；And D, E group Relative tumor proliferation rate remain close to 0%, effect always clearly, temporary nothing Recurrence trend.During testing, suppression ratio D, the E group of positive controls Relative tumor proliferation rate is in time Evening, 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 is 90.11 ± 2.76%, and 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 can be used for having larger carcinogenic risk except can be applied to breast cancer treatment, even Nonmalignant tumor removal completely, 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.
2. the non-knurl hyperplasia of mammary gland (non-struma is big) treatment
The proliferation of mammary gland is a kind of non-tumour benign proliferative enlargement, and clinical manifestation is with different degrees of breast pain It is characterized with lump in breast.Zoopery is summarized as follows：
Animal used：SPF grades of non-pregnant rat, all females of sex, the weight of animals is 150-180g during packet.
Animal models and is grouped：By above-mentioned rat muscle injection oestradiol benzoate 0.5mg/kg, 1 times/day, Continuous 20 days.Then intramuscular injection progesterone 5mg/kg, 1 times/day, continuous 5 days.After modeling terminates, take at random 6 rats are put to death, and take second pair of breast biopsy, see lobule of mammary gland and acinus showed increased, ductal epithelium increase, Cell growth is enlivened, and mammary gland is in secretor state, represents modeling success.Animal is entered using the softwares of PEMS 3.2 The random district's groups of row, are divided into blank control group (end modeling animal) and modeling successfully with the following group：Positive controls, Negative control group (A groups), drug research group.
Administration：The packet same day starts administration.The injected material of negative control group is 9% ethanol.Positive control is RUZENGNING PIAN, gavages, and administration number of times is 27 times, and administration frequency is for once a day, dosage is each 1g/kg. Each drug research group (B, C, D, E, F group) is injected in lump, and administration number of times is 5 times, and administration frequency is Per once two days.
Observation, measurement and analysis：
1) general states observation：Observation 1 time daily, self administration of medication starts to experiment the 28th day, observation index Or content includes but is not limited to animal administration part, appearance sign, general behavior activity, the state of mind, death Situations such as and other Novel presentations.
2) body weight determinations：Determined 1 time before administration and administration phase is determined weekly 2 times.
3) food rations are determined：Administration phase is determined weekly 1 time.
4) breast local form：Respectively at the 1st, 3,5,7,11,16,21,27 days after administration with trip Mark slide calliper rule accurate measurement second pair or so two papilla diameters (being designated as D) of rat.
It is with respect to papilla diameter computing formula：RTD=Dt/D0, D in formula0It is packet administration measurement for the first time Gained papilla diameter, DtPapilla diameter during to measure each time.
It is with respect to nipple Magnification computing formula：T/C (%)=TRTD/CRTD× 100, T in formulaRTDIt is the positive Control group or drug research group RTD, CRTDIt is negative control group RTD.
The evaluating drug effect standard of drug research group is：T/C (%)>50 for inactive, T/C (%)≤50 (but >And Analysis of variance P compared with negative control group 25%)<0.05 for active, T/C (%)≤25 and through variance Analysis P compared with negative control group<0.05 is have preferred activity.
5) breast tissues pathology changes：Put to death animal within second day after last dose, win rat second to mammary gland, Paraffin section, in the change of light Microscopic observation breast tissue pathology form after HE dyeing.By observe leaflet and The form of acinus, is integrated to each group rat breast tissue pathology, and 4 grades are divided into by pathological proliferation degree：Breast Glandular lobule not hyperplasia, body of gland quantity is few, and acinus is not expanded, and remembers 0 point；Lobule of mammary gland is individual without obvious hyperplasia Other acinus has slight hyperplasia, but without expansion, remembers 1 point；Lobule of mammary gland major part hyperplasia, part acinus substantially expands , remember 2 points；The obvious hyperplasia of lobule of mammary gland, acinus is in extreme expansion state, and glandular epithelium is in flat, There is substantial amounts of secretion in acinus and in conduit, remember 3 points；Mammary gland alveolus, conduit, the pathological proliferation of leaflet Substantially, 4 points are remembered.
The evaluating drug effect standard of drug research group is：Pathology integration >=3 for inactive, 1<Pathology is integrated<3 and pass through Variance analysis P compared with negative control group<0.05 is active, and pathology integrates≤1 and Analysis of variance and feminine gender Control group compares P<0.05 is have preferred activity.
The different degrees of appearance fur after modeling of animal is owed gloss, depilation, apocleisis, easily enrages etc. symptom. The all tumor-bearing mice body weight of drug research group and food ration show no obvious abnormalities change compared with negative control group, poor Different not statistically significant (P>0.05).
The packet same day, animal teat enlargement is upright, congested obvious, and there is yellow-white point on indivedual visible nipple surfaces Secretion is remained.Within whole experimental period, negative control group papilla diameter shows a rising trend；From experiment the 7th day, Each drug research group papilla diameter is reduced compared with negative control group in addition to B groups, and difference is statistically significant (P<0.05).Until the 19th day from testing the 10th day, each drug research group increases with respect to nipple beyond B groups Big rate (T/C) is respectively less than 50%；From testing the 19th day, C, D group are respectively less than 20% with respect to nipple Magnification, E groups are slow with respect to the decline of nipple Magnification.During testing, positive controls are with respect to nipple Magnification Suppression ratio C, D, E groups evening, significantly larger in data (be more than 30%) in time.After administration 20 days, the pathology integration≤1 of C, D, E group, and with significant difference (P<0.05) positive controls Pathology integration is more than 2.
More than during each experiment, each seminar has no animal dead, average weight and negative control group body weight Change represents medicine under experimental conditions without obvious toxic reaction close to (being dropped by less than 10%).Seminar with Positive controls compare, and general status are similar to.But transplantable tumor is tested positives control group average weight and is declined more (it is more than 20%) greatly, points out a stronger toxic reaction.
During the medication of target area, compared with negative control group, in seminar, especially in maximum concentration group There is the change of obvious color and luster most of injection site, even with myodegeneration, scope is more within 10mm. Further research explanation, these denaturation are that quinine dihydrochloride effect is caused.After stopping is administered, experimental group The musculature outward appearance of nude mice injection site is done step-by-step normalization.The nude mice for killing is cutd open after stopping to administration within 30 days Injection site carried out check pathological section, as a result show that test group is obviously reduced with the difference of negative control group, Tend to normal.
Also obtained with some other the quinolines material injections (such as the injection in table 1) prepared in embodiment 1 Obtain similar results.According to the above pharmacodynamic result and local side reaction result, the medicine group that the present invention is provided Optimum condition of the compound in mammary gland disease application is treated is as follows：Target area is administered；Quinolines material injection concentration is 0.05-0.5M, preferably 0.1-0.4M, more preferably 0.1-0.3M.Different quinolines material injections can have Slightly different preferred injection concentration scopes.For example, the injection containing quinine be 0.25-0.35M, containing a hydrochloric acid The injection of quinine is 0.2-0.35M, the injection containing quinine dihydrochloride is 0.2-0.3M, the note containing primaquine Agent is penetrated for 0.1-0.2M, the total concentration scope of quinolines composition injection are equal or slightly larger than containedization The concentration range of compound, etc..
Additionally, another battery of tests shows, pharmaceutical composition of the invention is under same concentration, and one larger Injection volume shows more preferable pharmacological effect.It is control dosage if Gross Target Volume is too big, can be right Target area carries out branch's treatment.Each administered volume of injection be necessary for part volume to be processed more than 70%, It is preferred that more than 110%.When the target block of target area is lump (such as tumour, non-struma block) and its is individual sufficiently small, Part volume to be processed is single lump volume.
Embodiment 4：Application in thyroid disease treatment
Clinically, most common thyroid disease is Thyroid Gland Swell and hyperthyroidism.In the present embodiment, to quinoline Class thing Quality Research is carried out first by a quinin hydrochloride.Each drug research group (B, C, D, E, F group) The preparation method of injection is identical with the preparation method of B2 in table 1, includes 2% phenmethylol but containing required The quinin hydrochloride of various concentrations one, injection concentration is respectively 0.025M, 0.05M, 0.15M, 0.25M, 0.4M, Ethanol content in injection is 5-9%.Injection dosage is less than mono- quinin hydrochlorides of 300mg/kg animals, injecting body Product is determined by injection dosage and concentration.
1. thyroid tumors treatment
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.Other experiment conditions and mammary gland in embodiment 3 Tumor therapy experiments are identical, and result of the test is also similar to, in general：
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).
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 (<25%).Test the 22nd day Rise, C, D group Relative tumor proliferation rate close to 15%, effect always clearly, temporarily without the trend of recurrence. During testing, each drug research group beyond the suppression ratio B groups of positive controls Relative tumor proliferation rate exists Evening, significantly larger in data (40% or so) on time.
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 90%). Positive controls tumour inhibiting rate is 61%.
Additionally, the present invention provide anti-knurl application in optimum condition under, pharmaceutical composition of the present invention is for connecing The effect for developing otherwise varied knurl body of the time of kind difference thus cancer cell, being also that inoculation time is shorter swells Knurl tumor killing effect is better, and it even can be used for having the non-of larger carcinogenic risk except can be applied to treatment of cancer The removal completely of malignant tumour, to prevent it to develop into malignant tumor.
2. goitre treatment
Simple goiter (simple goiter), also known as non-inflammatory goitre, is by non-inflammatory or tumprigenicity Reason causes thyroid gland compensatory enlargement caused by thyroxine dyssynthesis.By setting up iodine deficiency thyroid gland Swollen animal model, has carried out this research.
Experimental animal：From adult rat, male and female are not limited, body weight 100-200g.
Modeling and packet：During beginning, animal is raised with iodine-deficient forage (amount of iodine 50ng/kg weight in wet base feeds, master Contain corn 55%, soybean 15%, yeast 10%, milk powder 10%, low iodine mutton 9%, inorganic salts vitamin 1%).Progressively remove low iodine mutton and milk powder, rat is adapted to Low iodine diet without causing underfed hair It is raw.Rat is raised more than 3 months in the environment of iodine-deficient forage nursing.Be decreased obviously with urinating iodine, thyroid gland it is bright Aobvious enlargement is to model successfully.Random district's groups are carried out to animal using the softwares of PEMS 3.2, is divided into blank control group (end Modeling animal) and modeling it is successful with the following group:Positive controls, negative control group (A groups), drug research group, Every group 6.
Administration：The packet same day starts administration.Positive control is Potassiumiodate KIO3, gavaging, administration number of times is 27 times, administration frequency is for once a day, dosage is every time 0.4 μ g/kg.The injected material of negative control group is corresponding Solvent.Drug research group and negative control group are Thyroid Tumors and periphery injection, and administration number of times is 10 times, Administration frequency is per once two days.
Observation, measurement and analysis：
1) general states observation：Observation 1 time daily, self administration of medication starts to experiment the 22nd day, observation index Or content includes but is not limited to animal administration part, appearance sign, general behavior activity, the state of mind, death Situations such as and other Novel presentations.
2) body weight determinations：Determined 1 time before administration and administration phase is determined weekly 2 times.
3) food rations are determined：Administration phase is determined weekly 1 time.
4) local forms：Respectively at observation rat thyroid enlargement in the 1st, 6,11,16,21 days after administration.
5) .24h iodine discharge rate：24h urines and excrement are collected, 24h urine iodine and the discharge of excrement iodine is determined and calculate Amount.Urine iodide determination is using gentle acid nitrification arsenic-cerium reaction AAS.Excrement iodide determination uses sode ash method.
6) pathological examinations：Put to death within the 10th day after last dose animal, observation thyroid morphology, folliculus shape Fibr tissue form between state, epithelial cell form, leaflet.
In the whole test period, animal without death, without abnormal body temperature.
24h iodine discharge rate detection display before packet, modeling animal compares its iodine discharge rate with animal is not modeled Substantially reduce, the obvious enlargement of thyroid gland.From experiment the 11st day, each drug research group Thyroid Gland Swell with it is cloudy Property control group compared to being obviously improved, the reduction of iodine discharge rate is relative to ease up.Wherein, C, D, E group iodine discharge rate Reached unanimity (difference with blank control group<25%), effect is substantially than positive controls (with blank control group difference >35%) it is good.
In pathological examination, negative control group can be observed the obvious enlargement of thyroid gland, and folliculus is intensive, and epithelium is thin Born of the same parents' hypertrophy is in high column, it is seen that the cell mass rope of hyperplasia, interfollicular blood vessel showed increased, lumen distention Fibr tissue increases between hyperemia, leaflet.Can be observed to be obviously improved in each drug research group.Wherein, D, E groups are in thyroid size, folliculus form size, the aspect such as fibr tissue tends to just between epithelial cell, leaflet Often, hence it is evident that than positive controls closer to the state for not modeling rat.
3. Treatment of Hyperthyroidism
Hyperthyroidism (hyperthyroidism, HT) (abbreviation hyperthyroidism in the present invention), refers to by many The factor of kind causes internal thyroxine secretion excessive, causes to increase with system stimulants such as nerve, circulation, digestion With one group of general name of disease that hypermetabolism is main performance.Therefore, hyperthyroidism is a kind of clinical syndrome.
Experimental animal：Adult male SD rats, body weight 300-400g, 36-40 age in days.
Modeling and packet：Levothyrocine is dissolved in physiological saline, daily intraperitoneal injection, injection It is 50 μ g/100g body weight to measure, continuous injection 10 days.Then Serological testing and pathological examination are carried out, with Judge whether to model successfully.Random district's groups are carried out to animal using the softwares of PEMS 3.2, is divided into blank control group (end Modeling animal) and modeling it is successful with the following group：Positive controls, negative control group (A groups), drug research group, Every group 6.
Administration：The packet same day starts administration.Positive control is methimazol, is gavaged, and administration number of times is 27 times, Administration frequency is for once a day, dosage is each 2mg/kg.The injected material of negative control group is physiological saline. Drug research group and negative control group are thyroid body injection, and administration number of times is 10 times, and administration frequency is every Once two days.
Observation, measurement and analysis：
1) general states observation：Observe 1 time within every 2 days, self administration of medication starts to administration the 30th day, observation Index or content include but is not limited to animal administration part, appearance sign, general behavior activity, the state of mind, Dead situations such as and other Novel presentations.
2) body weight determinations：Determined 1 time before administration and administration phase is determined weekly 2 times.
3) food rations are determined：Administration phase is determined weekly 1 time.
4) Serological testing：Respectively at administration before and administration after the 30th day from animal to be checked extraction blood 5ml, Its serum T is determined using the method for exempting from (RIA) is put3、T4And thyrotropic hormone (TSH) concentration value.
5) pathological examinations：Respectively at administration before and administration after the 30th day by blood drawing after animal to be checked execution, Observation thyroid morphology, fibr tissue form between folliculus form, epithelial cell form, leaflet.Thyroid gland is visually Observation increases in diffusivity in various degree；Microscopic observation is in high column to follicular epithelium hyperplasia, and has small folliculus Formed；The many epithelial cells absorption vacuoles not of uniform size of folliculus periphery appearance, interstitial rich blood vessel, hyperemia, Lymphadenia.
Serological testing before packet shows that modeling animal compares its serum T with animal is not modeled3、T4Substantially Raise (raise more than 100%), TSH is then decreased obviously, and illustrates to model successfully.The blood of the 30th day upon administration During clear is checked, each drug research group serum T3、T4The T compared with negative control group3、T4Rising eases up, TSH Decline tends to improving.Wherein, the Serological testing result of C, D group is small with the blood serum values difference for not modeling animal In 25%, and the difference of positive controls is more than 35%.
In pathological examination, negative control group can be observed thyroid gland and increase in diffusivity, follicular epithelium hyperplasia In high column and there is small folliculus to be formed, folliculus periphery many epithelial cells not of uniform size occurs and absorbs vacuoles, Matter rich blood vessel, hyperemia, lymphadenia.Compared with negative control group, the thyroid gland of each drug research group Diffusivity has gone down, and the situation that follicular epithelium hyperplasia and small folliculus are formed is significantly reduced；What folliculus periphery occurred Epithelial cell absorbs vacuole, the number of interstitial blood vessel and congested and lymphadenia have been weakened.Each group first Shape gland diffusivity is in disappearance trend, and other pathological phenomenas relevant with hyperthyroidism have regression trend, close not model The state of animal, hence it is evident that better than positive controls.
During being tested more than, each seminar has no animal dead, average weight and negative control group changes of weight Close to (being dropped by less than 10%), represent medicine under experimental conditions without obvious toxic reaction.Seminar and the positive Control group compares, and general status are similar to.But transplantable tumor is tested positives control group average weight and is declined more greatly (greatly In 20%), a stronger toxic reaction is pointed out.
During the medication of target area, compared with negative control group, in seminar, especially in maximum concentration group There is the change of obvious color and luster most of injection site, even with myodegeneration, scope is more within 10mm. Further research explanation, these denaturation are that the collective effect of a quinin hydrochloride and ethanol is caused.Stop in administration After only, the musculature outward appearance of experimental group nude mice injection site is done step-by-step normalization.30 days after stopping to administration The injection site for cuing open the nude mice for killing has carried out check pathological section, as a result shows the difference of test group and negative control group It is not obviously reduced, tends to normal.
Also obtained with some other the quinolines material injections (such as the injection in table 1) prepared in embodiment 1 Obtain similar results.According to the above pharmacodynamic result and local side reaction result, the medicine group that the present invention is provided Optimum condition of the compound in thyroid disease application is treated is as follows：Target area is administered；Quinolines material injection concentration It is 0.05-0.5M, preferably 0.1-0.4M, more preferably 0.1-0.3M.Different quinolines material injections can be with There are slightly different preferred injection concentration scopes.For example, the injection containing quinine be 0.25-0.35M, containing a salt The injection of sour quinine be 0.2-0.35M, the injection containing quinine dihydrochloride be 0.2-0.3M, containing primaquine It is contained that injection is that 0.1-0.2M, the total concentration scope of quinolines composition injection are equal or slightly larger than The concentration range of compound, etc..
Additionally, another battery of tests shows, pharmaceutical composition of the invention is under same concentration, and one larger Injection volume shows more preferable pharmacological effect.It is control dosage if Gross Target Volume is too big, can be right Target area carries out branch's treatment.Each administered volume of injection be necessary for part volume to be processed more than 70%, It is preferred that more than 110%.When the target block of target area is lump (such as tumour, non-struma block) and its is individual sufficiently small, Part volume to be processed is single lump volume.

Claims (9)

1. a kind of pharmaceutical composition for treating thyroid gland, mammary gland disease, it includes quinolines material and medicine The upper acceptable excipient of thing, and be the formulation for being suitable for being administered through thyroid gland, mammary gland disease target area, wherein The quinolines material is selected from in the following group：Chloroquine, HCQ quinoline, ammonia first quinoline, amodiaquine, quinine, nitroquine, PIPERAQUINE, primaquine, pamaguine, pentane quinoline and its isomers and pharmaceutically acceptable salt, and through target The concentration of quinolines material is 0.05-0.5M in described pharmaceutical composition when area is administered.
2. the pharmaceutical composition described in claim 1, wherein the concentration of the quinolines material be 0.1-0.4M, Preferably 0.1-0.3M.
3. the pharmaceutical composition described in claim 1, wherein when target area is administered, the volume of the injection Not less than the Gross Target Volume for being intended to process 70%, preferably not less than 110%.
4. the pharmaceutical composition described in claim 1, wherein the quinolines material be quinine or primaquine its Isomers or pharmaceutically acceptable salt, such as a quinin hydrochloride, quinine dihydrochloride or their mixture.
5. one of claim 1-4 described pharmaceutical composition, wherein the pharmaceutically acceptable salt bag Include inorganic acid addition salt and organic acid addition salt, a such as hydrochloride, a hydrobromate, a hydriodate, two Hydrochloride, dihydrobromide, two hydriodates, disulfate, sulfate, phosphate, bicarbonate or carbon Hydrochlorate, ethyl carbonate salt, formates, acetate, Chinese holly edge hydrochlorate and tannate.
6. one of claim 1-4 described pharmaceutical composition, wherein the pharmaceutically acceptable figuration Agent includes water, the miscible organic solvent of water or their mixture, and the miscible organic solvent of water is for example 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, it is described hard Agent preferably includes one below kind or various：Lauromacrogol, sodium morrhuate, EO, poly- many cards Alcohol, bleomycin A5.
8. the pharmaceutical composition described in claim 1, wherein the disease includes enlargement, wherein the enlargement Including malignant tumour, non-malignant tumors, non-struma be big or non-knurl hyperplasia, tumour.
9. the pharmaceutical composition described in claim 1, wherein the disease includes hyperthyroidism.
CN201510923813.2A 2015-12-11 2015-12-11 Application, method and pharmaceutical composition of the quinolines material in treatment thyroid gland and mammary gland disease Pending CN106860459A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107929240A (en) * 2017-12-13 2018-04-20 桂林南药股份有限公司 Piperaquine phosphate oral liquid and preparation method thereof
WO2021068203A1 (en) * 2019-10-11 2021-04-15 中山大学附属第一医院 Use of chloroquine or derivative hydroxychloroquine
CN116444525A (en) * 2023-03-27 2023-07-18 成都苑东生物制药股份有限公司 Preparation method of ponatinib hydrochloride A crystal form

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1158250A (en) * 1996-12-04 1997-09-03 侯仁忠 hyperthyroidism injection and its prepn. method

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1158250A (en) * 1996-12-04 1997-09-03 侯仁忠 hyperthyroidism injection and its prepn. method

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
中华人民共和国药典编委会: "《中华人民共和国药典2005年版，二部》", 31 January 2005 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107929240A (en) * 2017-12-13 2018-04-20 桂林南药股份有限公司 Piperaquine phosphate oral liquid and preparation method thereof
WO2021068203A1 (en) * 2019-10-11 2021-04-15 中山大学附属第一医院 Use of chloroquine or derivative hydroxychloroquine
CN116444525A (en) * 2023-03-27 2023-07-18 成都苑东生物制药股份有限公司 Preparation method of ponatinib hydrochloride A crystal form

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