CN108938675A - A kind of caprophyl positioning transplantation method - Google Patents
A kind of caprophyl positioning transplantation method Download PDFInfo
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- CN108938675A CN108938675A CN201810866487.XA CN201810866487A CN108938675A CN 108938675 A CN108938675 A CN 108938675A CN 201810866487 A CN201810866487 A CN 201810866487A CN 108938675 A CN108938675 A CN 108938675A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/4841—Filling excipients; Inactive ingredients
- A61K9/4858—Organic compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
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Abstract
The present invention provides a kind of methods of caprophyl positioning transplanting, capsule is prepared into as content using caprophyl to be taken orally, the dissolution pH value of the capsule shells and the pH value of caprophyl transplanting target site are adapted, make the capsule in transplanting target site dissolution, by caprophyl positioning transplanting in target site.The different principle of the main different parts pH value according to human organ of the present invention, can make caprophyl capsule pinpoint release caprophyl according to not homologous transplantation target site, improve caprophyl in the transplanting efficiency of target transplantation site.
Description
Technical field
The present invention relates to microorganism pharmaceutical technology field, in particular to the positioning transplantation method of a kind of caprophyl.
Background technique
" caprophyl transplanting " (Fecal microbiota transplantation, FMT) refers to, will be in healthy human faecal mass
Functional flora is transplanted in patient's gastrointestinal tract, and new intestinal flora is rebuild, and realizes the treatment of enteron aisle and parenterally disease.Caprophyl
Transplant the crucial salvage treatment as refractory inflammatory bowel disease (Crohn disease, ulcerative colitis), the good effect played
Fruit.
Since use is viable bacteria for caprophyl transplanting, caprophyl needs to be transplanted to specific disease sites and is bred, ability
Enough play the effect for the treatment of intestines problem.Currently, standardization caprophyl transplanting, is in laboratory by Modern Scientific Instruments in Chinese human nature
Change ground separation and obtain highly purified flora, then the bacterium solution of quantization is infused into patient's enteral through scope or drainage tube.It is domestic
Caprophyl transplanting mode specifically includes that nasogastric tube, Nasal cavity intestinal tube, superior gastrointestinal endoscope (gastroscope), Sigmoidoscope are set pipe or retained and fills
Intestines.The shortcomings that these caprophyl transplanting modes be to operating technology require it is high, complicated for operation, inconvenient, patient suffering is not easily accepted by,
It is long to influence daily life quality, administration time.These caprophyl transplantation methods can not be connect by many clinical workerss and patient
By without the prospect for becoming routine treatment.
Summary of the invention
The purpose of the present invention is to provide a kind of methods of caprophyl positioning transplanting, can be according to the kind of disease using this method
Class is different, and caprophyl is transplanted to specific disease sites, easy to operate, no pain, patient compliance height.
Technical scheme is as follows:
The present invention provides a kind of methods of caprophyl positioning transplanting, are prepared into capsule as content using caprophyl and are taken orally,
The dissolution pH value of the capsule shells and the pH value of caprophyl transplanting target site are adapted, when the capsule moves to transplanting target
It is dissolved when position, the caprophyl positioning transplanting is in target site.
Preferably, the capsule shells dissolve in the range of pH value is 6~7, and small intestine is transplanted in caprophyl positioning.
Preferably, the capsule shells dissolve in the range of pH value is 7~8, and colon is transplanted in caprophyl positioning.
Preferably, the caprophyl is donor intestinal microecology integral transplanting.
Preferably, the extracting method of the caprophyl includes:
(A) donor excrement is mixed with sterile saline, stirs, obtains excrement slurry;
(B) be successively with aperture by the excrement slurry 2mm, 1mm, 0.5mm and 0.25mm strainer filtering, remove excrement
Residue and large particulate matter collect caprophyl filtrate;
(C) the caprophyl filtrate is centrifuged 3~5min with the rate of 1500~3000r/min, precipitating is resuspended in nothing
In bacterium physiological saline, caprophyl liquid is obtained.
It is furthermore preferred that the extraction of the caprophyl is completed in 20~40 minutes.
It is furthermore preferred that obtaining caprophyl dry powder after the caprophyl liquid is freeze-dried, the freeze-drying is complete within for 24 hours
At.
Preferably, the capsule is taken within production 30 days.
Preferably, also contain prebiotics in the content of the capsule.
It is furthermore preferred that the prebiotics be one of stachyose, galactooligosaccharide, Oligomeric manna sugar and xylo-oligosaccharide or
It is a variety of.
The prior art is compared, the invention has the following advantages that
The present invention provides a kind of methods of caprophyl positioning transplanting, are prepared into capsule as content using caprophyl and are taken orally,
The dissolution pH value of the capsule shells and the pH value of caprophyl transplanting target site are adapted, when the capsule moves to transplanting target
It is dissolved when position, the caprophyl positioning transplanting is in target site.The main different parts pH value according to human organ of the present invention is not
Same principle can make caprophyl capsule pinpoint release caprophyl according to not homologous transplantation target site.For different lesions type
Caprophyl positioning is transplanted to target transplantation site, improves caprophyl in the transplanting efficiency of target transplantation site by organ sites.
Specific embodiment
The present invention provides a kind of methods of caprophyl positioning transplanting, the different original of the different parts pH value according to human organ
Reason, is prepared into capsule as content using caprophyl and is taken orally, and the dissolution pH value of the capsule shells and the caprophyl transplant target portion
The pH value of position is adapted, the dissolution when the capsule moves to transplanting target site, and the caprophyl positioning transplanting is in target site.
The present invention is not particularly limited the type of caprophyl, using the conventional caprophyl type in this field.In this hair
In bright, preferably caprophyl be donor enteron aisle whole Tiny ecosystem.
The donor inclusion criteria of caprophyl of the present invention is preferred are as follows: (1) age 18-23 years old, health, unmarried men and women;
(2) rule of life, no bad habit;Antibiotic-free application history in (3) three months;(4) without enterogastric diseases;(5) serology and
The inspection of excrement infectious agent and culture are feminine gender;(6) intestinal flora type and diversity are good: probiotics type and flora
Diversity it is good (high-throughput intestinal flora 16srDNA genetic test).
The donor exclusion criteria of caprophyl of the present invention is preferred are as follows: (1) drug administration history: using antibiosis in three months
Element, cathartic, slimming drugs are taking immunosuppressor, chemotherapeutics person;(2) it viruses contact history: once or is in the recent period exposed to
Inhibition of HIV, hepatitis A virus, hepatitis B, hepatitis C virus person;(3) history of disease: psychotic disorder, pernicious obesity, constipation, glycosuria
Disease, inflammatory bowel disease, allergy, metabolic syndrome, hypoimmunity, autoimmune disease, enteron aisle related disease, confirmed fatigue
Syndrome, digestive system history of operation, gastrointestinal cancer or polyp person, atopic diseases, such as eczema, asthma and stomach and intestine
Road eosinophil related disorders etc.;(4) virology and aetology: Serologic detection human immunodeficiency virus (HIV), the mankind
Thermophilic T cell virus (HTLV), hepatitis A virus (HAV), hepatitis B (HBV), hepatitis C virus (HCV), cytomegalovirus, Epstein-Barr virus
(EBV), syphilis, quasi-colubriformis and Amoeba etc.;Fecal sample detects helicobacter pylori, Yersinia, campylobacter, will
Congratulate Pseudomonas, Salmonella, enteropathogenic E.Coli, rotavirus, adenovirus, astrovirus, toxoplasma and giardia lamblia etc.;
(5) high risk behavior history, drug abuse or forbidden drug have imprisonment history or epidemic-stricken area travel history using history, in the recent period.
Influence because of intestinal microflora vulnerable to diet is contributing the last week according to presetting for the flora that secures good health
Dietary reference table donor diet is adjusted, avoid taking in high sugared, high in fat and irritable food, eat fruits and vegetables more and be rich in
Dietary fiber food.
Caprophyl capsule should be administered at empty stomach in the receptor for receiving caprophyl transplanting (FMT);Constipation person should do enteron aisle taking the previous day
Cleaning, so that caprophyl capsule can preferably reach patient part on time.
The present invention is not particularly limited the existence form of caprophyl, and caprophyl of the present invention can be caprophyl liquid, can also be with
For caprophyl dry powder, as long as can guarantee that any caprophyl form of caprophyl microbial activity can be applied in the application.
The extracting method of caprophyl of the present invention preferably includes following steps:
(A) donor excrement is mixed with sterile saline, stirs, obtains excrement slurry;
(B) be successively with aperture by the excrement slurry 2mm, 1mm, 0.5mm and 0.25mm strainer filtering, remove excrement
Residue and large particulate matter collect caprophyl filtrate;
(C) the caprophyl filtrate is centrifuged 3~5min with the rate of 1500~3000r/min, precipitating is resuspended in nothing
In bacterium physiological saline, caprophyl liquid is obtained.
In the present invention, the sterile saline is preferably ice physiological saline, more preferably 4 DEG C stored refrigerated sterile lifes
Salt water is managed, the activity of flora is kept under low temperature.The mass volume ratio of the present invention preferably described excrement and the sterile saline
For 1g:(300~800) ml, more preferably 1g:(400~600) ml.In the present invention, preferably by donor excrement and sterile physiological salt
Water is stirred again after impregnating.It is preferred that the time impregnated is 1~2min, the time of stirring is preferably 1~2min.
In the present invention, the centrifugation rate is preferably 2000~2500r/min, and centrifugation time is preferably 4min.The present invention
The number of the centrifugation is preferably 1~3 time, the obtained precipitating of centrifugation mixed with sterile saline after centrifugation carry out again from
The heart.It is centrifuged obtained precipitating for the last time to be suspended in sterile saline again, obtains caprophyl liquid.It is preferably heavy in the present invention
Forming sediment with the volume ratio of sterile saline is 1:5~15, more preferably 1:8~11.
The extraction of caprophyl of the present invention is preferably completed in 20~40 minutes, more preferably 30min, keeps the steady of caprophyl
It is qualitative and active.Caprophyl liquid of the invention is immediately placed in -80 DEG C of low-temperature biological storage cabinets after completing and saves, when preservation
Between within 72 hours, guarantee the bioactivity of caprophyl.
Caprophyl of the present invention may be caprophyl dry powder.The caprophyl liquid cooling freeze-drying that above-mentioned technical proposal is extracted
Dry, preferably vacuum freeze drying obtains caprophyl dry powder.The temperature of the preferably described freeze-drying of the present invention is -50 DEG C~-53
DEG C, preferably -51 DEG C~-52 DEG C;The time of the freeze-drying preferably within for 24 hours, more preferably 18~for 24 hours, remove excrement
Moisture in bacterium solution.
Obtained caprophyl liquid or caprophyl dry powder is for making capsule.The present invention does not have special limit to the method for production capsule
It is fixed, using the conventional production method in this field.It is preferred that containing 1 × 10 in every caprophyl capsule being prepared10~2.88
×1010A bacterium, more preferably 2.88 × 1010It is a.The caprophyl capsule being prepared preferably is taken within 30 days, to guarantee
The bioactivity of caprophyl in capsule.
In the content of capsule of the present invention, other than containing caprophyl, it is also preferable to include prebiotics.The prebiotics
Preferably one of stachyose, galactooligosaccharide, Oligomeric manna sugar and xylo-oligosaccharide or a variety of ensure and improve the life of caprophyl
Object activity.In the present invention, the mass ratio of the preferably described caprophyl and the prebiotics is 5~10:2~5, more preferably 7~9:3~
4.The prebiotics mix addition preferably in the caprophyl liquid being prepared.
In the present invention, the capsule shells of the capsule can dissolve under specific pH value, the dissolution pH value of the capsule shells
It is adapted with the pH value of caprophyl transplanting target site, makes the capsule in transplanting target site dissolution, the caprophyl is determined
Displacement is planted in target site.The present invention is not particularly limited the source of capsule shells, can use commercial goods capsule shells, such as small
Intestines capsulae enterosolubilis, colon capsulae enterosolubilis etc..
The capsule shells dissolve in the range of pH value is 6~7 as a preferred technical solution,.Due to the pH value of small intestine
In the range of 6~7, meet the dissolution pH value of capsule shells.The oral capsule containing caprophyl, when capsule is moved to small intestine site
When, the caprophyl positioning release in capsule is transplanted to small intestine, caprophyl can be made to breed and play in small intestine by capsule shells dissolution
Its bioactivity.
Alternatively preferred technical solution, the capsule shells dissolve in the range of pH value is 7~8.Colon
PH value meets the dissolution pH value of capsule shells in the range of 7~8.The oral capsule containing caprophyl, when capsule is moved to colon portion
When position, the caprophyl positioning release in capsule is transplanted to colon, caprophyl can be made concurrent in small intestine breeding by capsule shells dissolution
Wave its bioactivity.
To make the objectives, technical solutions, and advantages of the present invention clearer, below with reference to embodiment to the present invention into
Row detailed description, but they cannot be interpreted as limiting the scope of the present invention.
Excrement donor used in the embodiment of the present invention is to meet donor inclusion criteria and donor exclusion criteria of the present invention
People.
Embodiment 1
(1) material is collected: fresh excreta is put into special feces collection bucket;
(2) bacterium solution dilutes: 100g excrement being added in 400ml ice physiological saline (4 DEG C stored refrigerated) and is impregnated 1 minute, is used
Blender stirs evenly, and about 1 minute;
(3) it filters: successively being filtered with stainless steel mesh, strainer mesh number is incremented by 2mm, 1mm, 0.5mm, 0.25mm step by step, goes
Except excrement residue and large particulate matter, caprophyl suspension is collected;
(4) eccentric cleaning: divide 3 times
1st time centrifugation: 5 minutes 2000 turns, remove supernatant, stay sediment (5ml), add physiological saline to 45ml, shake up;
2nd time centrifugation: 3 minutes 2000 turns, remove supernatant, stay sediment, add physiological saline to 45ml, shake up;
3rd time centrifugation: 3 minutes 2000 turns.Supernatant is removed, sediment is stayed, adds physiological saline to 20ml, while 3.7g weight is added
Amount shakes up than the stachyose and galactooligosaccharide that are 1:1;
(5) it is freeze-dried: by the caprophyl suspension packing freeze-drying bottle of purifying, being freezed at -20 DEG C 2 hours after being formed, on
It is freezed 24 hours under -50 DEG C of freeze drier, 20KPa, is prepared into dry caprophyl powder;
(6) it dispenses capsule: caprophyl powder being fitted into small intestine enteric capsule shells in Biohazard Safety Equipment and is prepared into capsule.Institute
Small intestine enteric capsule shells are stated, by weight include: 100 parts by weight of gelatin;3~8 parts by weight of water;8~15 weight of sodium alginate
Measure part;8~15 parts by weight of acrylic resin;20~45 parts by weight of n-butyl acetate;5~25 parts by weight of lauryl sodium sulfate;
0~8 parts by weight of colorant.Capsule shells can dissolve in the range of pH value 6~7, and every caprophyl capsule contains 2.88 × 1010A work
Property bacterium.
The oral caprophyl capsule being prepared, capsule dissolve under the action of small intestine intestinal juice, caprophyl positioning are migrated to small
Intestines position.
Disintegration time of 1 embodiment of table, the 1 caprophyl capsule in gastric juice and intestinal fluid
Embodiment 2
(1) material is collected: fresh excreta is put into special feces collection bucket;
(2) bacterium solution dilutes: 100g excrement being added in 500ml ice physiological saline (4 DEG C stored refrigerated) and is impregnated 1 minute, is used
Blender stirs evenly, and about 1 minute;
(3) it filters: successively being filtered with stainless steel mesh, strainer mesh number is incremented by 2mm, 1mm, 0.5mm, 0.25mm step by step, goes
Except excrement residue and large particulate matter, caprophyl suspension is collected;
(4) eccentric cleaning: divide 2 times
1st time centrifugation: 5 minutes 2500 turns, remove supernatant, stay sediment (5ml), add physiological saline to 45ml, shake up;
2nd centrifugation: 3 minutes 2500 turns, remove supernatant, stay sediment, add physiological saline to 20ml, while it is low that 3.7g is added
Poly- mannose, shakes up;
(5) it is freeze-dried: by the caprophyl suspension packing freeze-drying bottle of purifying, being freezed at -20 DEG C 2 hours after being formed, on
It is freezed 20 hours under -53 DEG C of freeze drier, 20KPa, is prepared into dry caprophyl powder;
(6) it dispenses capsule: caprophyl powder being fitted into colon enteric capsule shell in Biohazard Safety Equipment and is prepared into capsule, the glue
Softgel shell system gelatin adds empty hard capsule made of auxiliary material and suitable enteric material, is positioned at ileocecus and the disintegration of colon front end
The capsulae enterosolubilis of drug release can dissolve in the range of pH value 7~8, and product executes " Chinese Pharmacopoeia " standard, capsule model specification
There is No. 0-3, it is optional to have various colors.Every caprophyl capsule contains 2.88 × 1010A active bacteria.
The oral caprophyl capsule being prepared, capsule dissolve under the action of colon intestinal juice, and caprophyl positioning is migrated to knot
Intestines position.
Disintegration time of 2 embodiment of table, the 2 caprophyl capsule in gastric juice, intestinal fluid and colonic fluid
It can be illustrated by table 1 and 2, caprophyl capsule of the invention can position transplanting in small intestine and colon site, improve
Drug bioavailability reduces taking dose, avoids drug directly irritating and liver first-pass effect to gastrointestinal tract, reduces drug
Hepatotoxicity improves positioning transplantation effect.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of method of caprophyl positioning transplanting, which is characterized in that capsule is prepared into as content using caprophyl and is taken orally, it is described
The dissolution pH value of capsule shells and the pH value of caprophyl transplanting target site are adapted, when the capsule moves to transplanting target site
When dissolve, caprophyl positioning transplanting is in target site.
2. the method according to claim 1, wherein the capsule shells pH value be 6~7 in the range of dissolve,
Small intestine is transplanted in caprophyl positioning.
3. the method according to claim 1, wherein the capsule shells pH value be 7~8 in the range of dissolve,
Colon is transplanted in caprophyl positioning.
4. the method according to claim 1, wherein the caprophyl is donor enteron aisle entirety Tiny ecosystem.
5. the method according to claim 1, wherein the extracting method of the caprophyl includes:
(A) donor excrement is mixed with sterile saline, stirs, obtains excrement slurry;
(B) be successively with aperture by the excrement slurry 2mm, 1mm, 0.5mm and 0.25mm strainer filtering, remove excrement residue
And large particulate matter, collect caprophyl filtrate;
(C) the caprophyl filtrate is centrifuged 3~5min with the rate of 1500~3000r/min, precipitating is resuspended in sterile life
It manages in salt water, obtains caprophyl liquid.
6. according to the method described in claim 5, it is characterized in that, the extraction of the caprophyl is completed in 20~40 minutes.
7. according to the method described in claim 5, it is characterized in that, obtain caprophyl dry powder after the caprophyl liquid is freeze-dried,
The freeze-drying is completed within for 24 hours.
8. the method according to claim 1, wherein the capsule is taken within production 30 days.
9. the method according to claim 1, wherein also containing prebiotics in the content of the capsule.
10. according to the method described in claim 9, it is characterized in that, the prebiotics be stachyose, it is galactooligosaccharide, oligomeric
One of mannose and xylo-oligosaccharide are a variety of.
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CN201810866487.XA CN108938675B (en) | 2018-08-01 | 2018-08-01 | Method for positioning and transplanting fecal fungi |
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CN110178787A (en) * | 2019-05-08 | 2019-08-30 | 河南中医药大学 | A method of self-closing disease rat model is established with caprophyl grafting |
CN111888381A (en) * | 2020-09-18 | 2020-11-06 | 广西细微生物科技有限责任公司 | Intestinal microorganism transplanting process |
CN113209039A (en) * | 2021-04-22 | 2021-08-06 | 重庆晶云生物科技有限公司 | Enterobacter capsule and preparation method thereof |
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Cited By (4)
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CN110178787A (en) * | 2019-05-08 | 2019-08-30 | 河南中医药大学 | A method of self-closing disease rat model is established with caprophyl grafting |
CN110178787B (en) * | 2019-05-08 | 2022-03-22 | 河南中医药大学 | Method for establishing autism rat model by using coprophilous fungi transplantation method |
CN111888381A (en) * | 2020-09-18 | 2020-11-06 | 广西细微生物科技有限责任公司 | Intestinal microorganism transplanting process |
CN113209039A (en) * | 2021-04-22 | 2021-08-06 | 重庆晶云生物科技有限公司 | Enterobacter capsule and preparation method thereof |
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