CN102302496B - Applications of isoquinoline-type alkaloid and derivatives thereof in preparing medicaments for preventing or treating cervical cancer - Google Patents
Applications of isoquinoline-type alkaloid and derivatives thereof in preparing medicaments for preventing or treating cervical cancer Download PDFInfo
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Abstract
The invention relates to isoquinoline-type alkaloid for preventing and treating cervical cancer, belonging to the fields of preparation technique of natural medicament extractives and antitumor medicines. The invention also relates to applications of bulbocapnine, isocorydine, isoboldine, N-methyl hernovine, dicranostigma alkaloid or pharmaceutically acceptable salts or esters thereof in the preparation of medicaments for preventing and treating cervical cancer.
Description
Technical field
The present invention relates to a kind of isoquinoline alkaloid and derivant thereof, and uses thereof, natural drug extract and pharmaceutical applications technical field belonged to.
Background technology
Cervical cancer is that the whole world is only second to the second largest malignant tumor that breast carcinoma causes women's death, and its characteristics are the grade malignancy height, cures rate variance.WHO investigation in 2006 shows that the whole world has 490,000 examples to increase the cervical cancer patient newly every year, death surpasses 270,000, wherein have 85% to be found in developing country, the serious harm women's health, and Epidemiological study shows that sickness rate age of onset in rising trend is tending towards rejuvenation in recent years.Clinical treatment means for cervical cancer have surgical resection at present, and medicine chemotherapy and radiotherapy three big main body means in addition, also have emerging Biotherapeutics technology.But above several technology often is difficult to reach the long periods for the treatment of effect owing to exist in the treatment or the shortcomings such as bad complication after the treatment.Still lack effective treatment means at present.
Summary of the invention
The technical problem to be solved in the present invention has provided a kind of isoquinoline alkaloid and derivant thereof as prevention and treatment cervical cancer.
In order to solve the problems of the technologies described above, the invention provides following technical scheme:
Formula I or II chemical compound the application in prevention and treatment cervical cancer medication preparation of receptible salt or ester-formin;
Ⅰ
R
1Be H ,-OH ,-OCH
3,-O-C
1-C
8Alkyl ,-NH
2,-NO
2,-COOH ,-COOCH
3,-COOCH
2CH
3,-COO-C
1-C
8Alkyl ,-COO-fluoroalkyl, C
1-C
8Alkyl, C
3-C
6Cycloalkyl, C
1-C
8Carboxylic acid, C
1-C
8Carboxylate, 1-3 independent free halogen/HSCH
2-, CH
3SCH
2CH
2-;
R
2Be H ,-OH ,-OCH
3,-O-C
1-C
8Alkyl ,-NH
2,-NO
2,-COOH ,-COOCH
3,-COOCH
2CH
3,-COO-C
1-C
8Alkyl ,-COO-fluoroalkyl, C
1-C
8Alkyl, C
3-C
6Cycloalkyl, C
1-C
8Carboxylic acid, C
1-C
8Carboxylate, 1-3 independent free halogen/HSCH
2-, CH
3SCH
2CH
2-;
R
3Be H ,-OH ,-OCH
3,-O-C
1-C
8Alkyl ,-NH
2,-NO
2,-COOH ,-COOCH
3,-COOCH
2CH
3,-COO-C
1-C
8Alkyl ,-COO-fluoroalkyl, C
1-C
8Alkyl, C
3-C
6Cycloalkyl, C
1-C
8Carboxylic acid, C
1-C
8Carboxylate, 1-3 independent free halogen/HSCH
2-, CH
3SCH
2CH
2-;
R
4Be H ,-OH ,-OCH
3,-O-C
1-C
8Alkyl ,-NH
2,-NO
2,-COOH ,-COOCH
3,-COOCH
2CH
3,-COO-C
1-C
8Alkyl ,-COO-fluoroalkyl, C
1-C
8Alkyl, C
3-C
6Cycloalkyl, C
1-C
8Carboxylic acid, C
1-C
8Carboxylate, 1-3 independent free halogen/HSCH
2-, CH
3SCH
2CH
2-;
Ⅱ
R
1Be H ,-OH ,-OCH
3,-O-C
1-C
8Alkyl ,-NH
2,-NO
2,-COOH ,-COOCH
3,-COOCH
2CH
3,-COO-C
1-C
8Alkyl ,-COO-fluoroalkyl, C
1-C
8Alkyl, C
3-C
6Cycloalkyl, C
1-C
8Carboxylic acid, C
1-C
8Carboxylate, 1-3 independent free halogen/HSCH
2-, CH
3SCH
2CH
2-;
R
2Be H ,-OH ,-OCH
3,-O-C
1-C
8Alkyl ,-NH
2,-NO
2,-COOH ,-COOCH
3,-COOCH
2CH
3,-COO-C
1-C
8Alkyl ,-COO-fluoroalkyl, C
1-C
8Alkyl, C
3-C
6Cycloalkyl, C
1-C
8Carboxylic acid, C
1-C
8Carboxylate, 1-3 independent free halogen/HSCH
2-, CH
3SCH
2CH
2-;
R
3Be H ,-OH ,-OCH
3,-O-C
1-C
8Alkyl ,-NH
2,-NO
2,-COOH ,-COOCH
3,-COOCH
2CH
3,-COO-C
1-C
8Alkyl ,-COO-fluoroalkyl, C
1-C
8Alkyl, C
3-C
6Cycloalkyl, C
1-C
8Carboxylic acid, C
1-C
8Carboxylate, 1-3 independent free halogen/HSCH
2-, CH
3SCH
2CH
2-;
R
4Be H ,-OH ,-OCH
3,-O-C
1-C
8Alkyl ,-NH
2,-NO
2,-COOH ,-COOCH
3,-COOCH
2CH
3,-COO-C
1-C
8Alkyl ,-COO-fluoroalkyl, C
1-C
8Alkyl, C
3-C
6Cycloalkyl, C
1-C
8Carboxylic acid, C
1-C
8Carboxylate, 1-3 independent free halogen/HSCH
2-, CH
3SCH
2CH
2-.
Preferably, corydaline, isocorydine, isoboldine, N-methyl hernovine, tinea alba premium alkali or the acceptable salt of their medicine or the ester-formin application in prevention and treatment cervical cancer medication preparation.
A kind of pharmaceutical composition, said composition comprise each the acceptable salt of chemical compound or ester-formin and pharmaceutically-acceptable excipients or the carrier of claim 1-2 that contains the medicine effective dose.
The preferred salt form of chemical compound comprises the acceptable inorganic and formed salt of organic acid of those and medicine known in the art herein.The salt form that uses mineral acid to make comprises treatment upward acceptable halate and sulfate, for example hydrochlorate, maleate, hydrobromate, sulfate, carbonate or bicarbonate.These salt forms can use alkali compounds and the methods known in the art preparation of formula I or II.
The ester-formin of The compounds of this invention comprises straight chained alkyl ester or the branched alkyl ester that contains 3 or 6 carbon atoms or the benzyl ester with 1-6 carbon atom, comprises methyl, ethyl, propyl group, butyl, 2-methyl-propyl and 1,1-dimethyl ethyl ester.Input alkyl used herein comprises the carbochain of straight chain and side chain.Preferably, C1-C3 perfluoroalkyl substituent group is-CF3;-O-C1-C3 perfluoroalkyl substituent group is-OCF3 and-S-C1-C3 perfluoroalkyl substituent group is-SCF3.
Chemical compound of the present invention is the cervical cancer cell inhibitor, therefore is used for treating, suppress, prevent or prevent those to relate to the process that the pathology of development takes place cervical cancer at mammal, preferred human body.Therefore, The compounds of this invention is used for the treatment of or prophylactic treatment atypical hyperplasia pathological changes, cancer in situ, early invasive carcinoma, invasive squamous carcinoma, adenocarcinoma.
Chemical compound of the present invention also is used for prevention and treats the merging complication that other cause because of cervical cancer.
Because of the mechanism of action difference, The compounds of this invention can be united use with other antitumor drug and ancillary drug thereof.
The compounds of this invention can the enhancing body autoimmune function.The compounds of this invention can be used for improving the treatment of immunity of organisms aspect.
Treating, suppress, prevent or prevent the method for the listed every kind of disease of different animals of this paper is a part of the present invention.Every kind of method comprises The compounds of this invention or the acceptable salt of its medicine or ester-formin and the various pharmaceutical carrier excipient form of the mammal dose therapeutically effective that needs treatment.
The present invention also comprises compound structure formula I related among the present invention, the drug regimen that II is relevant.These combinations contain The compounds of this invention or the acceptable salt of its medicine or the ester-formin of dose therapeutically effective separately, perhaps with one or more pharmaceutical carriers or excipient phase mixed form.The medicine of chemical compound or treatment effective dose refer to that the amount of described chemical compound can fully suppress to need hyperfibrinolysis in the mammal for the treatment of herein, thereby provide improvement to described symptom to a great extent, and then prevent, suppress or limit the pathologic basis of described disease or symptom.
The employed exact dose of The compounds of this invention depends on symptom and the factors such as the order of severity, medication and employed excipient that comprise institute's administration object, the disease for the treatment of.Can give described chemical compound, its salt or ester-formin and contain pharmaceutical carrier and the excipient form of described chemical compound, salt or ester-formin by any conventional route.Form of medication comprises oral administration, intravenously administrable, percutaneous drug delivery, through intestinal canal administration, topical administration.Institute's administered compound can be dissociate or with suitable drug salts or the form of ester.The compounds of this invention can work in any mode with the fibrinolysin contact when treatment hyperfibrinolysis and hemorrhage.They can be used with any usual manner of administration that is applicable to, perhaps use as independent therapeutic dosage forms or with the combining form for the treatment of preparation.They can be used separately, but preferably use with suitable pharmaceutical carrier or excipient.Pharmaceutical carrier is selected according to selected route of administration, and can make is solid, liquid or solid and mixtures of liquids.
Solid comprises powder, tablet, capsule and nanometer formulation.Solid carrier can be one or more materials that can be used as flavoring agent, lubricant, solubilizing agent, suspending agent, binding agent or tablet disintegrant.In powder, described carrier is pulverizing solid, it and active component powder mixes.In tablet, active component with have essential adhesion characteristic carrier with suitable mixed and be pressed into required shape and size.Gelatine capsule contains active component and dust carrier, and example is known lactose, starch, cellulose derivative, magnesium stearate and stearic acid etc.Similarly, can use the diluent tabletting.Tablet and capsule can be made slow releasing preparation, discharge medicine continuously to continue a few hours.Tabletting can be with sweet tablet or film coating, with cover undesirable flavor make and make label and air broadcast from, perhaps carry out enteric coating with selectivity disintegrate in gastrointestinal tract.The liquid oral dosage form can contain coloring agent and correctives to increase patient's compliance.Suitable solid carrier is magnesium carbonate, magnesium stearate, Pulvis Talci, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, hydroxy methocel, sodium carboxymethyl cellulose, low melt wax, cocoa butter etc.Film-coat, become capsule material, nano material also can use with The compounds of this invention and salt thereof or ester-formin, term " compositions " refers to comprise active component and becomes capsule material as prescription, is with or without other carriers.
The sterile liquid compositions comprises solution, suspension, emulsion, slurry agent, tincture and spirit.Chemical compound of the present invention dissolves in or is suspended in the medicine acceptable carrier, for example sterilized water for injection.When the chemical compound sufficiently soluble, they can directly be dissolved in the normal saline of use or inapplicable appropriate organic solvent such as Polyethylene Glycol.If desired, chemical compound can be dispersed in starch solution, sodium carboxymethyl cellulose solution or the suitable cyclodextrin aqueous solution.Be the composition of liquid medicine of sterile solution or suspension, can pass through muscle, subcutaneous, Intradermal, abdominal cavity, intravenous injection and use.In many cases, can use forms of liquid compositions to replace the Peroral solid dosage form administering mode.
The unit dosage forms of the described chemical compound of preferred preparation carries out the standard administration.Unit dose can be mixed with packing powder, phial or ampoule, and be preferably capsule or tablet form.The daily dose of reactive compound can be according to the frequency of the other drug of the character of the pharmacodynamic properties of concrete medicine, route of administration, patient's body weight, age and sex, health status, morbid state and the order of severity, treatment simultaneously, treatment and by making corresponding adjustment to blood substance concentration analysis and patient's recovery situation and to the reaction for the treatment of.The daily dose of active component is 0.1 to 100 mg/kg, and preferred daily dose is about 1.0 to about 10 mg/kg.The combination dosage form that is suitable for administration comprises about l mg to about 10 mg active component/units.In these pharmaceutical compositions, the common content of active component accounts for the 0.5-95 % of composition total weight.Active component can be with solid dosage forms for example capsule, tablet and form of powder, can also be with the sterile liquid formulations administration.
Chemical compound of the present invention can pass through following experimental technique, determines that The compounds of this invention suppresses the ability of cervical cancer cell propagation:
1. extract and separate
Get dry Herba Dicranostigmatis Leptopodi (5.0 kg) and pulverize, with 95% ethanol extraction 3 times (each 7 days), decompression and solvent recovery gets extractum 200.0 g.Extractum is suspended in 2000 ml, 2% H
2SO
4In the aqueous solution, chloroform extraction part FA (12 g) successively, aqueous ph value is with ammonia modulation 9, chloroform extraction, extract FB(25 g), FB mixes sample with silica gel 25g, 500 g silica gel upper props, chloroform/methanol/triethylamine (10:1:0.1 is to 2:1:0.1) gradient elution, flow velocity, 10 ml/min, thin layer chromatography detects, and merges same composition, get 4 components, FB1(5000 ml, 4.0 g, 100:1), FB2(4000 ml, 4.0 g, 50:1), FB3(5000 ml, 2.0 g, 25:1) with FB4(3000 ml, 3.5 g, 5:1); FB1 is with 40g silica gel, and chloroform/methanol/triethylamine 20:1:0.1,10:1:0.1,8:1:0.1,6:1:0.1 are mobile phase, and the upper prop eluting obtains different isoquinoline alkaloid chemical compounds respectively.
2. MTT experiment
Principle: the isoquinoline alkaloid isocorydine that utilizes MTT experiment to detect variable concentrations carries 18HPV to Hela(), Siha(carries 16HPV), C33A(does not have HPV to be infected) influence of three kinds of cervical cancer cell proliferation activities.
Every hole with 5000 with the cell kind in 96 orifice plates, behind the growth 24h, give the isocorydine of variable concentrations, cultivate 24h respectively, behind 48h and the 72h, every hole adds MTT 20 μ l (final concentration is 5 mg/L), after cell is continued to cultivate 4h, liquid in the exhaustion hole, add 150 μ l DMSO (Sigma company), behind the horizontal oscillator tube vibration 10min, with Multiskan MS ELISA reader(Labsysterms, Helsinki Finland) detects absorbance (OD value) at the 490nm place.Experiment repeats 3 times.
The result: the MTT experiment that isocorydine suppresses human cervical carcinoma cell shows, as shown in Figure 1 and Figure 2, for the hela cell, shows the obvious suppression effect; For the Siha cell, show similar inhibitory action; And the C33A cell is that performance is the most responsive in three kinds of cervical cancer tumer lines, and inhibition is the strongest.Isocorydine shows the obvious depression effect to cervical cancer cell on the whole, and this effect of passing in time will continue.In addition, there is dependence in the concentration of the survival ability of cell and isocorydine.To have statistical significance (P ﹤ 0.05) by the one factor analysis of variance proof.
As shown in Figure 3, three kinds of cells give the isocorydine effect 24h of 1000 mg/L respectively, and 48h shows behind the 72h for isoquinoline alkaloid perception difference, when 72h, not as 24h and 48h.C33A finds more responsive to isocorydine, and the difference between the survival rate average is used in 24h, 48h, and three time points of 72h use the Tukeys post hoc analysis in the one factor analysis of variance, and the difference between average has statistical significance (P ﹤ 0.05).
3. transmission electron microscope observing:
Principle: Electronic Speculum is to observe ultrastructural pathology to change the most direct means, can observe nucleus intuitively, the change of cytoplasmic fine structure.By transmission electron microscope observing variable concentrations isocorydine (750mg/L, 1500mg/L, 3000mg/L) to cervical cancer cell Hela, Siha, the Ultrastructural influence of C33A, and observing apoptosis corpusculum.
Step: cell is with 5 * 10
6Density planted into 75cm
2Culture bottle in, cultivate 24h after, add the isocorydine of variable concentrations, establish the blank group simultaneously, behind the cultivation 48h, use 0.25% trypsin digestion and cell respectively, centrifuge tube is collected, the centrifugal 15min of 2000 rmp washes one time with PBS again, abandons supernatant, slowly the glutaraldehyde fixative of adding 0.25% fixedly spends the night for 4 ℃, the series dehydration is soaked into and embedding, and ultrathin section and dyeing are observed.
The result: as shown in Figure 4, after the cultivation 48.Electronic Speculum result shows: in the Hela cell, and the necrosis of H1 small part cytolysis, heterochromatin increases in the parts of fine karyon, chromatin margination, or see chromatin agglomerate shrinkage in the kytoplasm, see a large amount of autophagic vacuoles in the part cell cytoplasm; H2 sees more apoptotic cells, and cell membrane is imperfect; H3 sees a spot of apoptotic cell, most of cytolysis, necrosis.In the Siha cell, S1 group nucleus changes not obvious, but sees a large amount of autophagic vacuoles in the Cytoplasm; The existing chromatinic variation of S2 cell: the nuclear heterochromatin increases, and chromatin margination, and chromatin agglomerate in the kytoplasm have kytoplasm to change simultaneously again, a large amount of viable apoptotic cells occurs; S3 changes the group with S2, and different is to dissolve the S3 phase, and downright bad is that cell quantity is organized more than S2; C1 organizes more cytolysis, necrosis, and nucleus does not have obvious variation, sees a large amount of autophagic vacuoles in the kytoplasm; C2 group, it is imperfect that C3 organizes most of cell membrane, see disintegrate in the kytoplasm with the chromatin fragment, a large amount of apoptotic cells occurs.
4. the hochest33258 fluorescent dye is observed:
Principle: by the hochest33258 fluorescent dye to the variable concentrations isocorydine (750mg/L, 1500mg/L, 3000mg/L) Zuo Yong cervical cancer cell Hela, Siha, the form of C33A is observed.
Step: with cell with 2.5 * 10
6The density kind advance in 24 orifice plates, cultivate 24h after, behind the irritation cell generation apoptosis, exhaust culture medium, add 0.5 ml fixative, 4 ℃ are spent the night fixingly, remove fixative, wash twice with PBS, each shaking table shakes 3 min, adds 0.5 ml Hoechest, 33258 dyeing liquors again, dyeing 5min, remove dyeing liquor, wash twice with PBS, each 3 min exhaust liquid, drip anti-fluorescent quenching mounting liquid on microscope slide, cover the coverslip that posts cell, allow cells contacting mounting liquid, about excitation wavelength 350 nm, observation of cell nuclear morphology under the fluorescence microscope about emission wavelength 460 nm, the result as shown in Figure 5.
5. flow cytometer observation of cell apoptosis:
Principle: by the apoptosis situation that flow cytometer can be observed tumor cell, judge that with this medicine is to the inhibitory action of tumor cell.
Step: with 1 * 10
5Density the cell kind is advanced 25cm
2In the culture bottle, behind the cultivation 24h, discard old culture fluid, (750mg/L, 1500mg/L 3000mg/L) intervene, and cultivate 24h respectively, 48h, 72h, collecting cell (number about 1～5 * 10 to add the isocorydine that contains different pharmaceutical concentration
6Individual/ml), the centrifugal 5min of 1000 r/min discards culture fluid, reuse PBS washing once, the centrifugal PBS that discards, 70% the ethanol that adds the ice pre-cooling is fixed, 40 ℃ are spent the night, the centrifugal fixative that discards, resuspended 5 min of PBS, 400 purpose screen filtrations once, centrifugal 5 min of 1000 r/min discard PBS, add the dyeing of 1 ml PI dye liquor, 40 ℃ of lucifuge 30 min, flow cytometer detects, and the result is as shown in Figure 7.
6. the TUNEL method detects apoptosis:
Principle: the TUNEL method is utilized the effect of TdT enzyme, and 3 of the segment DNA in the apoptotic cell '-OH free-end is carried out efficient specific marker FITC-dUTP, uses fluorescence microscope directly to observe then.To the FITC that mixes, also can detect by the antibody FITC antibody of peroxidase labelling simultaneously, and can use optical microscope to observe after the colour developing chemical reaction.
Step: tumor cell is planted into that 24 orifice plates carry out creep plate, cultivate and discard old culture medium after 24 hours, clean with PBS, the neutral formalin with 4% (PBS prepares PH=7.4) room temperature is 15-30 min fixedly, after using PBS to clean fixed cell 2 times then, use 0.3% H under the room temperature
2O
2Methanol is handled cell 15-30 min, its objective is sealing inwardness peroxidase, after using PBS to clean cell, re-use Permeabilisation Buffer 100 μ l and react 2-5min on ice, PBS cleans cell, the reactant liquor 50 μ l (TdT Enzyme 5 μ l+Labeling Safe Buffer 45 μ l) that prepare the ice-cold placement in back are in advance added on the microscope slide, reaction 60-90 min in 37 ℃ of couveuses, clean stopped reaction with PBS, use the Anti-FITC HRP Conjugate of 70 μ l behind 37 ℃ of reaction 30 min, reuse PBS cleans, and uses under the DAB.H2O2 reactant liquor room temperature and carries out 10-15 min chromogenic reaction.Clean stopped reaction with sterile purified water at last, reuse 3% C.I. 42590 (Methyl Green) dyeing is by observation by light microscope cell dyeing situation, as shown in Figure 6.
Description of drawings
Accompanying drawing is used to provide further understanding of the present invention, and constitutes the part of description, is used from explanation the present invention with embodiments of the invention one, is not construed as limiting the invention.In the accompanying drawings:
The normal blank group human cervical carcinoma cell Hela of Fig. 1, Siha, the common light microscopic morphological observation figure of C33A;
Fig. 2 variable concentrations isocorydine alkali treatment Hela, Siha, the light microscopic morphological observation figure of C33A48h;
Shown in Figure 3, Hela, Siha, C33A give the isocorydine effect 24h of 1000 mg/L respectively, 48h, the perceptual statistical data figure that shows behind the 72h;
Fig. 4 be the variable concentrations isocorydine to cervical cancer cell Hela, Siha, the transmission electron microscope observing figure of C33A ultrastructural influence;
Fig. 5 be the variable concentrations isocorydine to cervical cancer cell Hela, Siha, the fluorescent dye of the hochest33258 of C33A influence is observed figure;
Fig. 6 is that TUNEL detects observation figure;
Fig. 7 is flow cytometer observation of cell observation of Apoptosis figure.
The specific embodiment
Below the preferred embodiments of the present invention are described, should be appreciated that preferred embodiment described herein only is used for description and interpretation the present invention, and be not used in restriction the present invention.
Example 1
Isocorydine suppresses the effect of tumor bearing nude mice cervical cancer
Medicine name: isocorydine
Chemical name:
1,2,10-trimethoxy-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinolin-11-ol
Medicines structure:
Dosage form: injection, injectable powder
Route of administration: quiet notes, quiet
Laboratory animal: female Balb/C mice
Inquire into isocorydine whether human cervical carcinoma Hela cell and Siha cell nude mouse tumor growth are existed inhibitory action, can cell death inducing, thus estimate isocorydine in treatment human cervical carcinoma's using value.
Experimental technique: female BALB/C nude mouse, 6 ~ 8 ages in week, body weight 20 ~ 30g, 0.25% trypsinization is Hela cell, the Siha cell of logarithmic growth, and the medium centrifugal washing of reuse serum-free 2 times is adjusted into 1x10 with cell density
7Individual cell/ml, and be suspended among the PBS standby.With iodophor disinfection nude inoculation position, Hela cell or Siha cell suspension that No. 6 injection needles (l ml syringe) are drawn 0.2 ml (contain 2 * 10 approximately
6Individual cell), cell inoculation is subcutaneous in the nape portion of nude mice.1. the nude mice that inoculates the Hela cell becomes (about 0.2 cm of diameter) after the tumor, is divided into 3 groups at random, 10 every group.The aseptic PBS of A group intratumor injection, 0.l ml/ time; B group intratumor injection isocorydine 0.25 mg/ time (concentration 2.5 mg/ml, 0.l ml/ time); C group intratumor injection isocorydine 0.5 mg/ concentration 0.5 mg/ml, 0.l ml/ time.1 week 3 times, continued for 3 weeks.2. inoculate siha cell nude mice and become (about 0.2 cm of diameter) after the tumor, be divided into 3 groups at random, 10 every group.Each organizes method of disposal with the Hela cell.Situations such as routine observation mice spirit, energy, reaction, diet and defecation, and gross tumor volume and the speed of growth.Packet transaction is after 3 weeks, and the dislocation of row cervical vertebra is put to death.
The result shows, treatment group nude mice drinking-water, feed are normal substantially, do not have untoward reaction such as suffer from diarrhoea, become thin, be movable slow, and nude mice of control group loss of appetite partly occurs, become thin, lazyly move, degradation situation under the respond.Each organizes intratumor injection position skin does not have redness, ulceration.Tumor tissues carries out perusal when peeling off from nude mice, is nodositas, and each organizes not of uniform size, and canescence, quality are hard partially, is difficult for separating with surrounding tissue.During the treatment, isocorydine treatment group and matched group compare, and tumor growth rate obviously slows down, and most for the treatment of group tumor is no longer grown up in medication mid-term, and what have diminishes, and the part tumor disappears substantially.Treat after 2l days, measurement discovery Hela cell and Silia cell therapy group tumor size and weight are obviously dwindled than matched group and are reduced, and difference has statistical significance.The tissue pathological slice of laboratory animal shows: the matched group oncocyte is intensive, arrangement disorder, and in the form of sheets, irregular nido distributes, cell has bright atypia, based on circle or ellipse, examine big engrain, chromatin becomes bulk, and obvious kernel is arranged, slurry is abundant, visible more karyokinesis picture, and a matter is less.It is more sparse that treatment group oncocyte is arranged, visible sheet non-structure necrotic area, and cell differentiation is than matched group oncocyte maturation, the nucleus engrain, pyknosis, less and regular, nucleocytoplasmic ratio reduces, and kernel reduces, visible apoptotic body.Illustrate that isocorydine improves cervical cancer cell strain differentiation capability, it is ripe that differentiation is tending towards, and proliferation activity reduces.Above animal example explanation: isocorydine can suppress the propagation of cervical cancer cell, has antitumor action.
Example 2
Capaurine alkali suppresses the effect of tumor bearing nude mice cervical cancer
Medicine name: capaurine alkali
Chemical name:
(6aS)-2,10,11-Trimethoxy-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinolin-1-ol
Medicines structure:
Dosage form: injection, injectable powder
Route of administration: quiet notes, quiet
Laboratory animal: female Balb/C mice
Inquire into capaurine alkali whether human cervical carcinoma Hela cell and Siha cell nude mouse tumor growth are existed inhibitory action, can cell death inducing, thus estimate the using value of capaurine alkali aspect the treatment human cervical carcinoma.
Experimental technique: female BALB/C nude mouse, 6 ~ 8 ages in week, body weight 20 ~ 30g, 0.25% trypsinization is Hela cell, the Siha cell of logarithmic growth, and the medium centrifugal washing of reuse serum-free 2 times is adjusted into 1x10 with cell density
7Individual cell/ml, and be suspended among the PBS standby.With iodophor disinfection nude inoculation position, Hela cell or Siha cell suspension that No. 6 injection needles (lml syringe) are drawn 0.2ml (contain 2 * 10 approximately
6Individual cell), cell inoculation is subcutaneous in the nape portion of nude mice.1. the nude mice that inoculates the Hela cell becomes (the about 0.2cm of diameter) after the tumor, is divided into 3 groups at random, 10 every group.The aseptic PBS of A group intratumor injection, 0.l ml/ time; B group intratumor injection capaurine alkali 0.25 mg/ time (concentration 2.5 mg/ml, 0.l ml/ time); C group intratumor injection capaurine alkali 0.5 mg/ concentration 0.5 mg/ml, 0.l ml/ time.1 week 3 times, continued for 3 weeks.2. inoculate siha cell nude mice and become (about 0.2 cm of diameter) after the tumor, be divided into 3 groups at random, 10 every group.Each organizes method of disposal with the Hela cell.Situations such as routine observation mice spirit, energy, reaction, diet and defecation, and gross tumor volume and the speed of growth.Packet transaction is after 3 weeks, and the dislocation of row cervical vertebra is put to death.
Experimental result shows, treatment group nude mice drinking-water, feed are normal substantially, do not have untoward reaction such as suffer from diarrhoea, become thin, be movable slow, and nude mice of control group loss of appetite partly occurs, become thin, lazyly move, degradation situation under the respond.Each organizes intratumor injection position skin does not have redness, ulceration.Tumor tissues carries out perusal when peeling off from nude mice, is nodositas, and each organizes not of uniform size, and canescence, quality are hard partially, is difficult for separating with surrounding tissue.During the treatment, capaurine alkali treatment group and matched group compare, and tumor growth rate obviously slows down, and most for the treatment of group tumor is no longer grown up in medication mid-term, and what have diminishes, and the part tumor disappears substantially.Treat after 2l days, measurement discovery Hela cell and Silia cell therapy group tumor size and weight are obviously dwindled than matched group and are reduced, and difference has statistical significance.Above animal example explanation: capaurine alkali can suppress the propagation of cervical cancer cell, has antitumor action.
Example 3
Isoboldine suppresses the effect of tumor bearing nude mice cervical cancer
Medicine name: isoboldine
Chemical name:
(6aS)-2,10-Dimethoxy-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-1,9-diol
Medicines structure:
Dosage form: injection, injectable powder
Route of administration: quiet notes, quiet
Laboratory animal: female Balb/C mice
Inquire into isoboldine whether human cervical carcinoma Hela cell and Siha cell nude mouse tumor growth are existed inhibitory action, can cell death inducing, thus estimate the using value of isoboldine aspect the treatment human cervical carcinoma.
Experimental technique: female BALB/C nude mouse, 6 ~ 8 ages in week, body weight 20 ~ 30g, 0.25% trypsinization is Hela cell, the Siha cell of logarithmic growth, and the medium centrifugal washing of reuse serum-free 2 times is adjusted into 1x10 with cell density
7Individual cell/ml, and be suspended among the PBS standby.With iodophor disinfection nude inoculation position, Hela cell or Siha cell suspension that No. 6 injection needles (l ml syringe) are drawn 0.2ml (contain 2 * 10 approximately
6Individual cell), cell inoculation is subcutaneous in the nape portion of nude mice.1. the nude mice that inoculates the Hela cell becomes (about 0.2 cm of diameter) after the tumor, is divided into 3 groups at random, 10 every group.The aseptic PBS of A group intratumor injection, 0.l ml/ time; B group intratumor injection isoboldine 0.25 mg/ time (concentration 2.5 mg/ml, 0.l ml/ time); C group intratumor injection isoboldine 0.5 mg/ concentration 0.5 mg/ml, 0.l ml/ time.1 week 3 times, continued for 3 weeks.2. inoculate siha cell nude mice and become (about 0.2 cm of diameter) after the tumor, be divided into 3 groups at random, 10 every group.Each organizes method of disposal with the Hela cell.Situations such as routine observation mice spirit, energy, reaction, diet and defecation, and gross tumor volume and the speed of growth.Packet transaction is after 3 weeks, and the dislocation of row cervical vertebra is put to death.
Experimental result shows, treatment group nude mice drinking-water, feed are normal substantially, do not have untoward reaction such as suffer from diarrhoea, become thin, be movable slow, and nude mice of control group loss of appetite partly occurs, become thin, lazyly move, degradation situation under the respond.Each organizes intratumor injection position skin does not have redness, ulceration.Tumor tissues carries out perusal when peeling off from nude mice, is nodositas, and each organizes not of uniform size, and canescence, quality are hard partially, is difficult for separating with surrounding tissue.During the treatment, isoboldine treatment group and matched group compare, and tumor growth rate obviously slows down, and most for the treatment of group tumor is no longer grown up in medication mid-term, and what have diminishes, and the part tumor disappears substantially.Treat after 2l days, measurement discovery Hela cell and Silia cell therapy group tumor size and weight are obviously dwindled than matched group and are reduced, and difference has statistical significance.Above animal example explanation: isoboldine can be by suppressing the propagation of cervical cancer cell.
It should be noted that at last: the above only is the preferred embodiments of the present invention, be not limited to the present invention, although with reference to previous embodiment the present invention is had been described in detail, for a person skilled in the art, it still can be made amendment to the technical scheme that aforementioned each embodiment puts down in writing, and perhaps part technical characterictic wherein is equal to replacement.Within the spirit and principles in the present invention all, any modification of doing, be equal to replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (4)
1. isocorydine, capaurine alkali or the isoboldine application in prevention and treatment cervical cancer medication preparation.
2. isocorydine, capaurine alkali, isoboldine or the acceptable salt of their the medicine application in prevention and treatment cervical cancer medication preparation.
3. application according to claim 2 is characterized in that: the acceptable salt of described medicine comprises the formed salt of the acceptable mineral acid of medicine.
4. application according to claim 3 is characterized in that: the formed salt of described mineral acid comprises hydrochlorate, hydrobromate, sulfate, carbonate, bicarbonate, maleate.
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| WO2015180482A1 (en) * | 2014-05-30 | 2015-12-03 | 资元堂生物科技股份有限公司 | Use for aporphine alkaloid derivative in preparation of medicament promoting ampk activity |
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| CN102631345B (en) * | 2012-03-28 | 2019-11-05 | 上海市肿瘤研究所 | Pharmaceutical composition and its application for reversing tumor cellular drug resistance |
| CN102657653B (en) * | 2012-03-28 | 2017-10-17 | 上海市肿瘤研究所 | A kind of agent of tumor stem cell selective killing and its application |
| CN104447552A (en) * | 2014-12-11 | 2015-03-25 | 上海壹志医药科技有限公司 | Pharmaceutical use of corydine derivative |
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| US6123943A (en) * | 1997-12-22 | 2000-09-26 | Kaken Shoyaku Co., Ltd. | NF-KB activity inhibitor |
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| US6123943A (en) * | 1997-12-22 | 2000-09-26 | Kaken Shoyaku Co., Ltd. | NF-KB activity inhibitor |
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| Title |
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| 秃疮花生物碱成分分析及药理作用研究进展;赵强等;《陇东学院学报》;20100331;第21卷(第2期);第53-57页 * |
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| WO2015180482A1 (en) * | 2014-05-30 | 2015-12-03 | 资元堂生物科技股份有限公司 | Use for aporphine alkaloid derivative in preparation of medicament promoting ampk activity |
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