CN100588429C - PH sensitive type anticancer pro-drug, preparation method and uses thereof - Google Patents
PH sensitive type anticancer pro-drug, preparation method and uses thereof Download PDFInfo
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- CN100588429C CN100588429C CN200810306463A CN200810306463A CN100588429C CN 100588429 C CN100588429 C CN 100588429C CN 200810306463 A CN200810306463 A CN 200810306463A CN 200810306463 A CN200810306463 A CN 200810306463A CN 100588429 C CN100588429 C CN 100588429C
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Abstract
The invention relates to a pH-sensitive anticancer prodrug and a preparation method as well as application thereof, which belongs to the field of medicine and chemical industry. The technical problemto be solved is to provide the pH-sensitive anticancer prodrug, and the anticancer prodrug has highest solubility when the pH value is between 4 and 6, can nearly not be dissolved when the pH value isless than 3 or more than 7, and almost does not release adriamycin during the circulation due to small solubility particularly when circulating in blood (the pH value is approximately equal to 7.4),thereby having small toxic side effect. The pH-sensitive anticancer prodrug is a conjugate obtained by coupling the adriamycin and pectin with a molecular weight of between 71,000 and 400,000 througha chemical bond. The pH-sensitive anticancer prodrug is obtained by the following steps: a pectin aqueous solution and an adriamycin aqueous solution are added with a dehydrating agent to react underthe condition that the pH value is between 6 and 8, and then are dialyzed and condensed.
Description
Technical field
The present invention relates to a kind of pH sensitivity type anticancer prodrug and its production and use, belong to field of medicine and chemical technology.
Background technology
In recent years, the macromole targeted anticancer medicine becomes the emphasis to the chemotherapeutics research of lymph metastasis, this class conjugate has bigger molecular volume, behind local injection, can not see through the very narrow endothelium gap of blood capillary and enter blood, and enter partial draining lymph node than the endothelium gap that is easier to by the lymphatic capillary broad.In addition, this class conjugate can also take regional nodes to by by the mode of macrophage phagocytic, under the effect of lymph node endoenzyme, is hydrolyzed and discharges active substance (cancer therapy drug), plays the effect of killing metastasis cancer cell in the lymph node.Thereby this class macromolecular carrier targeted drug is suitable for the treatment that lymphatic cancer shifts most.
Pectin is commonly used for tablet, microsphere, the clothing layer of microcapsule and liposome etc., preparation drop pill and granule, embedding liquid or solid medicine, play the uncomfortable stink of slow releasing function and masking agents, it is natural macromolecular polysaccharide polymer, extensively be present in the cell wall of plant, by the acid macromolecular polysaccharide (HyunjoKim that α-(1 → 4)-D-galactopyranose aldehydic acid unit forms, et al.International Journal of Pharmaceutics, 1998,161:149-159), its molecule constitutes a helicoidal structure by 3 units, and the pitch of its spiral is 1.34nm.In addition, also have acetyl group and the sugared side chain of other neutrality (many) in its structure, as existence such as rhamnose, galactose, arabinose xyloses; Pectin can be by strengthening mononuclear phagocyte system, and activating macrophage, T cell and B cell, NK cell and complement system promote cytokine secretion, strengthen raising host immune functions such as erythrocyte immune; Growth characteristics by changing the solid carcinoma cell membrane, influence signal pipeline in the solid carcinoma cell, anti-radical action, induce differentiation and apoptosis, the nucleic acid and the protein synthesis of inhibition solid carcinoma cell, influence the solid carcinoma cell ultrastructure, influence oncogene, antimutagenic effect, the vascularization of inhibition solid carcinoma bring into play direct antitumaous effect (Chinese Chinese medicine information magazine, 1999,5:64).Pectin is water-soluble under the condition of pH 6-8, but reduces with pH, and dissolubility will reduce.
Pectin-amycin conjugate is suitable for preparing lympha targeted medicine, hepatic targeting drug or lung targeted drug, but if pectin-amycin conjugate (pH ≈ 7.4) circulation time in blood, also no show cancer entity has discharged amycin, then can cause toxic and side effects, reduce the therapeutic effect of amycin simultaneously.
Summary of the invention
Technical problem to be solved by this invention provides a kind of pH sensitivity type anticancer prodrug, this anticancer prodrug is 4～6 o'clock dissolubility maximums at pH, and at other condition indissoluble, especially (pH ≈ 7.4) circulation time in blood, because dissolubility is little, in cyclic process, discharge amycin hardly, thereby toxic and side effects is little.
Technical scheme of the present invention: the pectin with good biocompatibility is carrier, directly by covalent bond with amycin and pectin coupling, this covalent bond energy is by the hydrolysis and slowly release cancer therapy drug gradually of intravital enzyme.
Specifically, it is to be 7.1 ten thousand～400,000 (mean molecule quantity 15-30 ten thousand by chemical bond with amycin and molecular weight, preferred 200,000) conjugate that pectin coupling obtains, wherein the weight proportion of pectin, amycin is: 1 part of pectin, 0.05～1 part of amycin; This anticancer prodrug is 4～6 o'clock dissolubility maximums at pH, and at pH less than 3 or almost insoluble greater than 7 o'clock.
Second technical problem to be solved by this invention provides the preparation method of above-mentioned pH sensitivity type anticancer prodrug, it be by pectin aqueous solution and amycin aqueous solution under the condition of pH 6-8, add dehydrant reaction back dialysis, concentrate and get.
Its building-up process is as follows:
A, be that 7.1 ten thousand～400,000 pectin is water-soluble with molecular weight, be adjusted to pH 6-8 with sodium hydroxide solution, and add dehydrant behind the amycin aqueous solution mixing, 20～80 ℃ of temperature controls stir 3～24h; Preferable reaction temperature is 50 ℃, and the response time is 8h;
B, transfer to bag filter, the water dialysis, dialysate is concentrated into to be done promptly.
Described dehydrant is N, N-dicyclohexylcarbodiimide (being called for short DCC), N-cyclohexyl-N '-dimethylamine propyl carbodiimide (being called for short CDC), 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (being called for short EDCHCl), preferred EDCHCl is as dehydrant.
Pectin is natural polymer, have excellent biological compatibility, its construction features makes it possess lympha targeted characteristic, after local injection, can enter into injection site draining lymph node on every side specifically, the carrier that can be used as the lympha targeted property new drug of design uses.Pectin is water-soluble under the condition of pH 6-8, but along with pH value reduces, dissolubility will reduce.
The pectin that different preparation methoies or preparation condition prepare is different with the sensitivity to pH value of amycin conjugate, the inventor has also prepared water miscible pectin-amycin conjugate (seeing comparative experimental example for details), its dissolubility has identical trend with pectin, reduces and reduces rapidly with pH.The dissolubility of pectin of the present invention-amycin conjugate reduces along with the reduction of pH increases afterwards earlier, and is 5 o'clock dissolubility maximums at pH, and is that 7.4 o'clock dissolubility are very little at pH, and pH is that 5 o'clock dissolubility is that pH is 7.4 o'clock 14 times.After this shows injection this pectin-amycin conjugate, (pH ≈ 7.4) circulation time in blood,, in cyclic process, discharge amycin hardly, thereby toxic and side effects is little because dissolubility is little.And when granule by cytophagy, enter lysosome (pH ≈ 5) after, dissolubility can increase, thus slow release anti-cancer medicine amycin, so this conjugate has good pH sensitivity.PH sensitivity type anticancer prodrug of the present invention can be used for the treatment of and suppress tumor lymphatic metastasis, treatment hepatocarcinoma or pulmonary carcinoma.Can pass through local injection administration or intravenously administrable, can be expelled in the solid carcinoma cancer, also can be expelled in the solid carcinoma draining lymph node on every side, conjugate enters behind the lymph node by huge has a liking for cytophagy, hydrolysis discharges cancer therapy drug under the effect of enzyme in lymph node, is used for the treatment of solid carcinoma (for example breast carcinoma, gastric cancer, esophageal carcinoma, incidence cancer, cervical cancer, endometrial carcinoma, colorectal cancer etc.) and draining lymph node thereof and shifts.
The present invention has following advantage compared with the prior art:
The synthetic pectin of the present invention-amycin conjugate has pH sensitivity, at pH is 4-6, and especially pH is the dissolubility that dissolubility is much higher than other condition in 5 the phosphate buffer, and is that 5 o'clock particle diameter is little at pH, narrow distribution can be engulfed by tumor cell preferably.
Description of drawings
Fig. 1 pectin of the present invention-amycin conjugate and the water soluble pectin-dissolubility of amycin conjugate in phosphate buffer
● the dissolubility of pectin of the present invention-amycin conjugate
The dissolubility of the pectin of ■ contrast test-amycin conjugate
Fig. 2 pectin of the present invention-release conditions of amycin conjugate in phosphate buffer
Fig. 3 pectin of the present invention-amycin conjugate is particle size distribution figure in 3 the phosphate buffer at pH
Fig. 4 pectin of the present invention-amycin conjugate is particle size distribution figure in 5 the phosphate buffer at pH
Fig. 5 pectin of the present invention-amycin conjugate is particle size distribution figure in 7.4 the phosphate buffer at pH
Fig. 6 melanoma lung metastasis model mice time-survivor curve figure (n=10).
◆ pectin of the present invention-amycin conjugate group; ■ doxorubicin hydrochloride group; ▲ matched group.
The specific embodiment
Below by the specific embodiment the present invention is further described, be limitation of the invention but should not be construed as.Those of ordinary skills can also make modification, replacement, the change of various ways according to technique scheme.All modification, replacement, changes of doing based on above-mentioned technological thought all belong to scope of the present invention.
Below by embodiment the present invention is further described.
The preparation of embodiment 1 pectin of the present invention-amycin conjugate
Take by weighing 1g pectin (molecular weight 7.1 ten thousand～400,000, average mark molecular weight 200,000), add 100ml water, behind the pectolysis, regulate pH to 6-8 with sodium hydroxide solution.The 0.5g doxorubicin hydrochloride is dissolved in 100ml water, transfers in the pectin solution, temperature control stirred 30 minutes at 50 ℃.Add 1g EDCHCl, control temperature at 50 ℃, stirring reaction 6.5 hours.Behind the time to go, transfer to molecular cut off and be 7000 bag filter, with secondary water dialysis 24 hours, dialysate was concentrated into dried, obtains red solid 0.9g.
The preparation of embodiment 2 pectin of the present invention-amycin conjugates
Take by weighing 1g pectin (molecular weight 7.1 ten thousand～400,000, average mark molecular weight 200,000), add 100ml water, behind the pectolysis, regulate pH to 6-8 with sodium hydroxide solution.The 0.5g doxorubicin hydrochloride is dissolved in 100ml water, transfers in the pectin solution, temperature control stirred 30 minutes at 50 ℃.Add 1g EDCHCl, control temperature at 50 ℃, stirring reaction 8 hours.Behind the time to go, transfer to molecular cut off and be 7000 bag filter, with secondary water dialysis 24 hours, dialysate was concentrated into dried, obtains red solid 1.1g.
The preparation of embodiment 3 pectin of the present invention-amycin conjugates
Take by weighing 1g pectin (molecular weight 7.1 ten thousand～400,000, average mark molecular weight 200,000), add 100ml water, behind the pectolysis, regulate pH to 6-8 with sodium hydroxide solution.The 0.5g doxorubicin hydrochloride is dissolved in 100ml water, transfers in the pectin solution, temperature control stirred 30 minutes at 30 ℃.Add 1g EDCHCl, control temperature at 30 ℃, stirring reaction 24 hours.Behind the time to go, transfer to molecular cut off and be 7000 bag filter, with secondary water dialysis 24 hours, dialysate was concentrated into dried, obtains red solid 1g.
The preparation of embodiment 4 pectin of the present invention-amycin conjugates
Take by weighing 1g pectin (molecular weight 7.1 ten thousand～400,000, average mark molecular weight 200,000), add 100ml water, behind the pectolysis, regulate pH to 6-8 with sodium hydroxide solution.The 0.5g doxorubicin hydrochloride is dissolved in 100ml water, transfers in the pectin solution, temperature control stirred 30 minutes at 40 ℃.Add 1g EDCHCl, control temperature at 40 ℃, stirring reaction 10 hours.Behind the time to go, transfer to molecular cut off and be 7000 bag filter, with secondary water dialysis 24 hours, dialysate was concentrated into dried, obtains red solid 0.8g.
The preparation of embodiment 5 slightly solubilities pectin-amycin conjugate
Take by weighing 1g pectin (molecular weight 7.1 ten thousand～400,000, average mark molecular weight 200,000), add 100ml water, behind the pectolysis, regulate pH to 6-8 with sodium hydroxide solution.The 0.5g doxorubicin hydrochloride is dissolved in 100ml water, transfers in the pectin solution, temperature control stirred 30 minutes at 60 ℃.Add 1g EDCHCl, control temperature at 60 ℃, stirring reaction 5 hours.Behind the time to go, transfer to molecular cut off and be 7000 bag filter, with secondary water dialysis 24 hours, dialysate was concentrated into dried, obtains red solid 0.9g.
The preparation of embodiment 6 pectin of the present invention-amycin conjugates
Take by weighing 1g pectin (molecular weight 7.1 ten thousand～400,000, average mark molecular weight 200,000), add 100ml water, behind the pectolysis, regulate pH to 6-8 with sodium hydroxide solution.The 0.5g doxorubicin hydrochloride is dissolved in 100ml water, transfers in the pectin solution, temperature control stirred 30 minutes at 70 ℃.Add 1g EDCHCl, control temperature at 70 ℃, stirring reaction 6 hours.Behind the time to go, transfer to molecular cut off and be 7000 bag filter, with secondary water dialysis 24 hours, dialysate was concentrated into dried, obtains red solid 1.1g.
The preparation of comparative example's 1 water soluble pectin-amycin conjugate
Take by weighing 1g pectin (molecular weight 7.1 ten thousand～400,000, average mark molecular weight 200,000), the mixed solution that adds 100ml dimethyl formamide and water (volume ratio is 1: 1), behind the pectolysis, regulate pH to 6-8 with sodium hydroxide solution, then the 0.5g doxorubicin hydrochloride is dissolved in the mixed solution of 100ml dimethyl formamide and water (volume ratio is 1: 1), add pectin solution, temperature control stirred 30 minutes at 50 ℃.Add 1g EEDQ, control temperature at 50 ℃, stirring reaction 8 hours.Behind the time to go, be concentrated to about 100ml, add 300ml ethanol, produce precipitation, centrifugal.Precipitation use water dissolution, transfers to molecular cut off and be 7000 bag filter, dialyses 24 hours with secondary water.After liquid concentrated in the bag filter, vacuum drying got water soluble pectin-amycin conjugate 0.9g.
The preparation of comparative example's 2 water soluble pectins-amycin conjugate
Take by weighing 1g pectin (molecular weight 7.1 ten thousand～400,000, average mark molecular weight 200,000), the mixed solution that adds 100ml dimethyl formamide and water (volume ratio is 1: 1), behind the pectolysis, regulate pH to 6-8 with sodium hydroxide solution, then the 0.5g doxorubicin hydrochloride is dissolved in the mixed solution of 100ml dimethyl formamide and water (volume ratio is 1: 1), add pectin solution, temperature control stirred 30 minutes at 70 ℃.Add 1g EEDQ, control temperature at 70 ℃, stirring reaction 6 hours.Behind the time to go, be concentrated to about 100ml, add 300ml ethanol, produce precipitation, centrifugal.Precipitation use water dissolution, transfers to molecular cut off and be 7000 bag filter, dialyses 24 hours with secondary water.After liquid concentrated in the bag filter, vacuum drying got water soluble pectin-amycin conjugate 1g.
The preparation of comparative example's 3 water soluble pectins-amycin conjugate
Take by weighing 1g pectin (molecular weight 7.1 ten thousand～400,000, average mark molecular weight 200,000), the mixed solution that adds 100ml dimethyl formamide and water (volume ratio is 1: 1), behind the pectolysis, regulate pH to 6-8 with sodium hydroxide solution, then the 0.5g doxorubicin hydrochloride is dissolved in the mixed solution of 100ml dimethyl formamide and water (volume ratio is 1: 1), add pectin solution, temperature control stirred 30 minutes at 40 ℃.Add 1g EEDQ, control temperature at 40 ℃, stirring reaction 24 hours.Behind the time to go, be concentrated to about 100ml, add 300ml ethanol, produce precipitation, centrifugal.Precipitation use water dissolution, transfers to molecular cut off and be 7000 bag filter, dialyses 24 hours with secondary water.After liquid concentrated in the bag filter, vacuum drying got water soluble pectin-amycin conjugate 0.8g.
The mensuration of test example 1 conjugate drug loading
1, the preparation of standard curve
Take by weighing the 10mg doxorubicin hydrochloride, be dissolved in the 100ml secondary water, pipette 1,2,3,4 respectively, 5ml in the 10ml color comparison tube, be diluted with water to scale.Get concentration and be respectively 9.37,18.74,28.11,37.48,46.85 μ g/mL titers, locate to measure absorbance in 480nm (the ultraviolet-uisible spectrophotometer spectral scan is determined).With concentration absorbance is returned, getting regression equation is A=0.0174C-0.0028 (R
2=0.9927)
2, the pectin of the present invention-preparation of amycin solution and the mensuration of drug loading
Take by weighing the slightly solubility pectin-amycin 10mg of embodiment 2 preparations, add the hydrochloric acid solution of 80ml pH=2, ultrasonic dissolution, standardize solution is measured absorbance to 100ml at 480nm then, the substitution regression equation, as calculated, drug loading is 19.8%.
3, the comparative example's pectin-preparation of amycin solution and the mensuration of drug loading
Take by weighing the water soluble pectin-amycin 10mg of comparative example's 1 preparation, add the 80ml deionized water, ultrasonic dissolution, standardize solution is measured absorbance to 100ml at 480nm then, the substitution regression equation, as calculated, drug loading is 18.9%.
The test example 2 pectin-solubility test of amycin in different pH phosphate buffers
Pectin-amycin the conjugate of embodiment 2 preparations and the pectin-amycin conjugate and the dissolubility of pectin in the phosphate buffer of different pH (3,4,5,6,7.4,8) of comparative example's 1 preparation have been examined or check.See Fig. 1.
The result shows, the dissolubility of pectin of the present invention-amycin conjugate reduces along with the reduction of pH increases afterwards earlier, and is 5 o'clock dissolubility maximums at pH, and is that 7.4 o'clock dissolubility are very little at pH, and pH is that 5 o'clock dissolubility is that pH is 7.4 o'clock 14 times.After this shows injection this pectin-amycin conjugate, (pH ≈ 7.4) circulation time in blood,, in cyclic process, discharge amycin hardly, thereby toxic and side effects is little because dissolubility is little.And when granule by cytophagy, enter lysosome (pH ≈ 5) after, dissolubility can increase, thus slow release anti-cancer medicine amycin, so this conjugate has good pH sensitivity.For water soluble pectin-amycin conjugate of comparative example, its dissolubility in phosphate buffer then reduces with pH and reduces rapidly.
The test example 3 pectin-release test of amycin in different pH phosphate buffers
Pectin-amycin conjugate of having examined or check the pectin-amycin conjugate of embodiment 2 preparations and comparative example's 1 preparation is a release conditions in 3,5 and 7.4 the phosphate buffer at pH.
The result shows that the pectin-amycin of comparative example's 1 preparation does not all discharge amycin in the phosphate buffer of three kinds of pH, and the pectin-amycin of embodiment 2 preparations reduces along with pH value, and the amount that discharges amycin in the identical time increases, and sees Fig. 2.
Test example 4 is measured the particle diameter of pectin-amycin in different pH phosphate buffers
Get the pectin-amycin conjugate of 20mg embodiment 2 preparation, placing 10ml pH respectively is 3,5 and 7.4 phosphate buffer, stirs 5 hours, and the centrifugal 15min of 2000rpm gets supernatant with nano particle size instrument mensuration particle diameter.The result shows: in pH was 3 phosphate buffer, pectin-amycin particle diameter mainly was distributed between the 3091-5560nm; In pH was 5 phosphate buffer, pectin-amycin particle diameter mainly was distributed between the 342-458nm; In pH was 7.4 phosphate buffer, pectin-amycin particle diameter mainly was distributed between the 396-825nm.Therefore, the particle diameter of this conjugate also has pH sensitivity.Pectin of the present invention-amycin conjugate is that particle size distribution figure sees Fig. 3,4,5 in 3,5 and 7.4 the phosphate buffer at pH.
After test example 5 pectin of the present invention-amycin conjugate whole body administration in the intravital distribution test of mice
1, is subjected to the reagent thing
A, take by weighing the 21.4mg doxorubicin hydrochloride, be dissolved in the 10ml normal saline, obtain containing the injection of amycin 2mg/ml.
B, take by weighing pectin-amycin 101mg (being equivalent to contain amycin 20mg) of embodiment 2, add 100mg poloxamer 188, behind the mixing, the carboxymethylcellulose sodium solution that adds several 0.5%, grind 60min with grinding platinum, transfer to the volumetric flask of 10ml, the carboxymethylcellulose sodium solution standardize solution with 0.5% obtains the suspensoid injectio that doxorubicin concentration is 2mg/ml.
C, take by weighing the pectin-amycin 106mg (being equivalent to contain amycin 20mg) of comparative example 1 preparation, use physiological saline solution, standardize solution obtains containing the injection of amycin 2mg/ml to 10ml.
All injections were sterilized with gamma-radiation before doing zoopery.
2, animal
Kunming mouse, male and female half and half, body weight 18～22g is available from Sichuan University's West China medical college animal center.
3, method
3.1 medication
Choose 50 mices, be divided into 5 groups at random, 10 every group.Only give tail vein injection a, b, c and normal saline 0.1ml/ respectively.
3.2 sample collecting and processing
2 hours excision eyeballs are got blood after the administration, put to death mice.Get an amount of heart, liver, spleen, lung and renal tissue with homogenate (60% ethanol that contains 0.3mol/L hydrochloric acid) by weight volume ratio be made into tissue homogenate at 1: 9.The centrifugal 10min of 3000rpm after the homogenate.Get the 0.1ml supernatant and carry out fluoroscopic examination.The centrifugal 10min of blood sample 3000rpm.Get the 0.1ml supernatant and carry out fluoroscopic examination.
4, result
The fluorophotometric value of amycin sees Table 1 in the different tissues sample.
The fluorophotometric value (n=10) of amycin in 2 hours different tissues samples after table 1 administration
| Project | Heart | Liver | Spleen | Lung | Kidney | Blood |
| a | 353.07 ±65.27 | 303.48 ±27.83 | 248.9 ±47.51 | 280.78 ±25.89 | 273.42 ±30.14 | 40.42 ±2.519 |
| b | 297.41 ±35.26 | 483.15 ±69.38 | 277.13 ±20.14 | 570.81 ±42.13 | 367.43 ±63.52 | 44.301 ±0.213 |
| b1 | 243.5 ±16.69 | 887.99 ±103.62 | 248.15 ±26.93 | 488.89 ±25.71 | 211.89 ±15.69 | 44.297 ±0.568 |
| c | 291.86 ±30.84 | 569.88 ±75.63 | 224.67 ±35.87 | 523.58 ±42.35 | 283.4 ±59.87 | 43.504 ±2.634 |
| d | 166.63 ±22.16 | 177.62 ±34.11 | 71.03 ±12.04 | 51.45 ±18.20 | 132.19 ±14.24 | 37.79 ±1.635 |
Annotate: a. amycin normal saline group; B. pectin of the present invention-amycin normal saline; B1 pectin of the present invention-amycin suspensoid injectio group; C. comparative example's water soluble pectin-amycin group; D. normal saline group.
5, conclusion
Different doxorubicin formulations are not quite similar in the intravital distribution of mice.Slightly solubility pectin-amycin of pectin-amycin of the present invention and comparative example all has the trend of enrichment in liver and lungs, distinguishes to some extent with the preparation difference, but concentration in the blood all.Illustrate that pectin-amycin has certain become liver property and lung targeted characteristic.
The experimentation of test anti-mouse lung metastatic tumor of example 6 pectin-amycin of the present invention and pulmonary carcinoma
1. main material
Take doxorubicin hydrochloride injection and pectin of the present invention-amycin injection in the test example 5.
2. animal and cell strain
The C57BL/6J mice, female, 18～22g, 4～6 ages in week are available from Sichuan University's Experimental Animal Center.
Mice Bearing Lewis Lung Cancer cell line (LL/2) is purchased in ATCC (American Type Culture Collection), and this laboratory is protected and planted.
3. lung cancer model preparation and treatment
The lung cancer model preparation: collect the LL/2 cell of exponential phase, the centrifugal 3min of 1500rpm, cell precipitation washs 1 time with the culture medium of serum-free, antibiotic-free, and adjusting cell concentration with serum-free, antibiotic-free culture medium after the counting cells quantity is 1 * 10
7/ ml.Every mice is in tail vein injection 1 * 10
6Individual (0.1ml) tumor cell, 3 days begin treatments after the kind tumor.
Treatment: divide three groups, matched group (normal saline), amycin group (dosage 10mg/kg), slightly solubility pectin-amycin group (being equivalent to amycin dosage 10mg/kg), tail vein injection, volume injected 100ml, per 5 days 1 time, totally 3 ~ 4 times.
4. result
Different pharmaceutical is seen Fig. 6 to the influence of pulmonary carcinoma tumor-bearing mice life span; Each is organized medicine and pulmonary carcinoma tumor-bearing mice pulmonary is shifted what the influence of joint footing sees Table 2.
Table 2 different pharmaceutical shifts what the influence (n=10) of joint footing to pulmonary carcinoma tumor-bearing mice pulmonary
In a word, pectin-amycin of the present invention is compared the life span that all can prolong the pulmonary carcinoma tumor-bearing mice with the doxorubicin hydrochloride group with matched group, suppresses the growth of cancer joint knot in pulmonary.Pectin of the present invention-amycin effect is more obvious.
Claims (9)
1, a kind of pH sensitivity type anticancer prodrug, it is characterized in that: it is to be the conjugate that 7.1 ten thousand～400,000 pectin coupling obtains by chemical bond with amycin and molecular weight, this conjugate be by pectin aqueous solution and amycin aqueous solution under the condition of pH 6-8, add dehydrant reaction back dialysis, concentrate and get; Wherein the weight proportion of pectin, amycin is: 1 part of pectin, 0.05～1 part of amycin.
2, pH sensitivity type anticancer prodrug according to claim 1, it is characterized in that: described dehydrant is N, N-dicyclohexylcarbodiimide, N-cyclohexyl-N '-dimethylamine propyl carbodiimide or 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride.
3, the preparation method of pH sensitivity type anticancer prodrug is characterized in that its building-up process is as follows:
A, be that 7.1 ten thousand～400,000 pectin is water-soluble with molecular weight, be adjusted to pH 6-8, and add dehydrant behind the amycin aqueous solution mixing, 20～80 ℃ of temperature controls stir 3～24h;
B, transfer to bag filter, with the dialysis of secondary water, dialysate is concentrated into to be done promptly.
4, the method for preparing the pH sensitivity type anticancer prodrug according to claim 3, it is characterized in that: described dehydrant is N, N-dicyclohexylcarbodiimide, N-cyclohexyl-N '-dimethylamine propyl carbodiimide or 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride.
5, the method for preparing the pH sensitivity type anticancer prodrug according to claim 3 is characterized in that: reaction temperature is 50 ℃, and the response time is 8h.
6, claim 1 or the 2 described pH sensitivity type anticancer prodrugs purposes in the medicine of preparation treatment cancer.
7, claim 1 or the 2 described pH sensitivity type anticancer prodrugs purposes in preparation treatment lung cancer drugs.
8, pH sensitivity type anticancer pharmaceutical preparation is characterized in that: it is to add pharmaceutically acceptable complementary composition by claim 1 or 2 described pH sensitivity type anticancer prodrugs to be prepared from.
9, pH sensitivity type anticancer according to claim 8 pharmaceutical preparation is characterized in that: described anti-cancer drug preparation is local injection administration or intravenous administration formulation.
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| CN101721714B (en) * | 2009-12-18 | 2011-12-07 | 重庆莱美药业股份有限公司 | Passive targeting antitumor prodrug of solid tumor and preparation method thereof |
| CN102232932B (en) * | 2010-04-27 | 2013-06-05 | 重庆莱美药业股份有限公司 | Pectin-adriamycin coniuncate lyophilized preparation, and preparation method thereof |
| CN105311642B (en) * | 2014-05-28 | 2018-11-02 | 重庆莱美药业股份有限公司 | A kind of synthesis technology of pectin anticancer prodrug |
| CN109157662B (en) * | 2018-06-06 | 2021-07-20 | 北京大学 | Human serum albumin-adriamycin cross-linked substance nano-particles and application thereof |
| US11596643B2 (en) * | 2019-06-18 | 2023-03-07 | Sichuan Yingrui Pharmaceutical Technology Company | Pectin-adriamycin conjugate as well as preparation method and use thereof |
| CN110152013B (en) * | 2019-06-18 | 2022-04-26 | 四川瀛瑞医药科技有限公司 | Pectin-adriamycin conjugate and preparation method and application thereof |
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