CN1057192C

CN1057192C - Method for preparing preservation liquid for various kinds of living organs and the prepns. thereof

Info

Publication number: CN1057192C
Application number: CN97106600A
Authority: CN
Inventors: 郑军华; 闵志廉
Original assignee: Shanghai Changzheng Hospital
Current assignee: Shanghai Changzheng Hospital
Priority date: 1997-09-10
Filing date: 1997-09-10
Publication date: 2000-10-11
Anticipated expiration: 2017-09-10
Also published as: CN1176738A

Abstract

The present invention provides a method for preparing improved multiple-organ preserving liquid, and a product thereof, which is characterized in that the preserving liquid is added with low-molecular dextran-40, sucrose and calcium ion antagonist, original adding components are removed, and the pH value is regulated to be weakly-alkaline. The method of the present invention is easy to control. The multiple-organ preserving liquid prepared and produced by the method of the present invention has the advantages of low cost, good preserving effect and convenient storage, transportation and use, and can satisfy the requirements of the field of organ transplantation, which is increasingly developed. By the success of a low-temperature preserving test for animals, and low-temperature preservation and clinical transplantation for partial organs of a human body, the product of the present invention is the effective multiple-organ preserving liquid.

Description

A kind of method of preparing multi-organ preservation solution

The present invention relates to the preservation technology of medical domain human body, animal body or its part, is a kind of method and goods thereof of preparing new multi-organ preservation solution.

The China organ transplant General Board Xia Suisheng of association professor points out in " kidney transplant of expectation China obtains some suggestions of the bigger progress " commentary of " Chinese organ transplant magazine " the 15th the 4th phase of volume in October, 1994: " the initiative success of UW liquid (by Univ Wisconsin-Madison USA; University ofWiscosin; a kind of multi-organ preservation solution that professor Belzer etc. succeeded in developing in 1987; be the most successful in the world up to now multi-organ preservation solution; be with historically new significance on the organ transplant history; be called for short UW liquid), prolong the cold holding time of donor organ significantly (can reach 72 hours) for kidney, make preservation liquid enter a new period, extensive use in the world, domestic also have indivedual units to begin to use UW liquid in liver transfer operation, expectation China not only just introduces, but utilize it to preserve principle, the UW type solution of developing the homemade multiple improvement of China comes, and uses to apply." owing to lack and to be similar to UW liquid or to be better than the so long-acting multi-organ preservation solution of UW liquid; in a sense; restriction and obstacle the development of other the big organ transplant work except that kidney transplant, so develop a kind of new multi-organ preservation solution and be significant.

The object of the present invention is to provide a kind of method and goods thereof of preparing new more efficiently multi-organ preservation solution, further reduced cost, increase preservation effect, conveniently store, transport and use, can satisfy the needs of growing organ transplant circle.

The object of the present invention is achieved like this is at first being analyzed on the basis of UW formula of liquid comprehensively meticulously, and in conjunction with the new development of Organ Preservation, is improved.Process for preparation is in distilled water, add lactobionic acid, potassium hydroxide, sodium hydroxide, adenosine, allopurinol, potassium dihydrogen phosphate, magnesium sulfate, sucrose, reduced glutathione, isoptin, dexamethasone, dextran-40 by quantitatively strict and condition order, carry out pH value and capacity adjustment again.There are 5 points in the place of change: 1. substitute HES (E.I.Du Pont Company's specialities) with D-40-40 (molecular weight is 40.000), further to reduce cost, increase preservation effect, especially aspect microcirculation; 2. with cane sugar substitution kapok sugar; 3. remove adding ingredient in the UW liquid, as insulin, heparin, penicillin, Batricn (trimethoprim (TMP)); 4. add calcium ion antagonist-isoptin; 5. the pH value further is adjusted into inclined to one side alkali (7.45 ± 0.10).

The present invention has following advantage and positive effect: a kind of method of preparing new more efficiently multi-organ preservation solution provided by the present invention is easy to grasp, preparation and production that prescription that those of ordinary skill can provide by this method and step are carried out this multi-organ preservation solution.Multi-organ preservation solution with this method preparation and production is a kind of new more efficiently multi-organ preservation solution.Be characterized in: reduced cost, increased preservation effect, made things convenient for storage, transportation and use, can satisfy the needs of growing organ transplant circle.Through animal organ's's (kidney, liver, heart, lungs, pancreas, small intestine etc.) low temperature being preserved experiment and trying out in the low temperature preservation of part human organ and the success of clinical transplantation, confirm a kind of new more efficiently multi-organ preservation solution of the present invention, low temperature preservation effect to kidney, liver, heart, lungs, pancreas, small intestine etc. is analogous to external UW liquid substantially, and partial results is better than UW liquid, approximately can preserve kidney 72 hours, liver 30 hours, heart 18-24 hour, pancreas 48 hours etc.Its usage is: available simple hypothermia perfusion method or consecutive low temperature machine method for filling are preserved each organ.

Below further set forth concrete prescription of the present invention and implementation step.

The inventor recommends following prescription, its composition content (/L) scope is respectively:

1, lactobionic acid (Lactobionate Acid) 99～101mmol

2, potassium dihydrogen phosphate (KH ₂PO ₄) 24～26mmol

3, magnesium sulfate (MgSO ₄) 4～6mmol

4, adenosine (Adenosine) 4～6mmol

5, glutathione reduced form (Glutathione) 2～4mmol

6, dexamethasone (Dexame thasone) 8mg

7, allopurinol (Allopurinol) 0.5～1.5mmol

8, sodium hydroxide 5N (NaOH 50mmol/L) 4～6ml

9, potassium hydroxide 5N (KOH 5mmol/L) 19～21ml

10, sucrose (Sucrose) 50～70mmol

11, dextran-40 (Dextran-40) 40-60g

12, isoptin (Isoptin) 20mg

pH：7.45+0.10：Permeate?pressure：310±10mOsm/L

(osmotic pressure: 310 ± 10mOsm/L)

Concrete steps are as follows:

List process for preparation by the 1000ml amount below:

1. distilled water: the distilled water that in 1 liter container, adds 400ml earlier.It is 20-24 ℃ generally in room temperature.

2. lactobionic acid: add the 35.83g lactobionic acid, stir become until liquid color clear.

3. potassium hydroxide: add 20ml 5N potassium hydroxide, and stirred 5 minutes.

4. sodium hydroxide: add 5.0ml 5N sodium hydroxide, and stirred 5 minutes.

5. adenosine: add adenosine 1.34g, stir become until liquid color clear.

6. allopurinol: add the 0.136g allopurinol, stir become until liquid color clear.

7. potassium dihydrogen phosphate: add the 3.4g potassium dihydrogen phosphate, stir become until liquid color clear.

8. magnesium sulfate: add 0.60g magnesium sulfate, stir become until liquid color clear.

9. sucrose: add 20.538g sucrose, stir become until liquid color clear.

10. glutathione: add the 0.92g glutathione, stir become until liquid color clear.

11. isoptin: add the isoptin of 20ml, it is clear that stirring becomes until liquid color.

12. dexamethasone: add the 8mg dexamethasone, it is clear that stirring becomes until liquid color.

13. dextran-40: add dextran-40 500ml (10%), heat and dissolve.

14.pH the adjusting of value: as conditioning agent, pH is adjusted to 7.45 ± 0.10 with the sodium hydroxide of 5N.

15. capacity adjustment: add distilled water to 1000ml.And stirred 5 minutes.

Above-mentioned process for preparation is all prepared under strict aseptic condition.

This multi-organ preservation solution adopts the aseptic filtration sterilization.

This multi-organ preservation solution needs to store under 0-4 ℃ of temperature.

This multi-organ preservation solution is the 2-3 month the 0-4 ℃ of term of validity.

Prepare the chemical reagent source of this multi-organ preservation solution:

1, lactobionic acid (Lactobionate Acid)

Aldrich?Chemical?Co.，Milwaukee，WI，#15.351-6.97％

FW：358.30 mP113～118℃

[α]D20+25°(C＝10，H2，24hrs)

Beil.31，415，Merck?Index?11，5219

NMR?2(1)，460B?FT-IR?1(1)，5250，Disp.A

2, adenosine (Adenosine)

Sigma?A?9251

C ₁₀H ₁₃N ₅O ₄

FW：267.2

3, glutathione reduced form (Glutathione.Reduced from)

Sigma?G?4251

C ₁₀H ₁₂N ₃O ₆S

FW：307.3

4, allopurinol (Allopurinol) 0.5-1.5mmol

Sigma?A?8003

4-Hydroxypyrazolo[3，4-d]-pyrimidine；HPP

C ₅H ₄N ₄O

FW：136.1

5, isoptin (Isoptin)

Verapamil?hydrochloride

composition：

2ml?of?injection?solution?contain?5?mg?of?verapamil?hydrochloride

Knoll?AG.D6700?Ludwigshafen.?Germany

6, sucrose (Sucrose)

Shanghai reagent one factory

C ₁₂H ₁₂O ₁₁

FW：342.30

Specific rotatory power [α] 20/D+66.40～+ 66.60

7, dextran-40 claims glucan (Dextran-40) again

Chemical reagent station, Shanghai packing factory

FW：40.000

8, sodium hydroxide (Sodium Hydroxide)

NaOH

Reagent four factories in Shanghai supervise

FW：40.00

NaOH content is no less than 96.0%

Na ₂CO ₃Content is not more than 1.5%

9, potassium dihydrogen phosphate (Potassium Phosphate Monobasic)

Shanghai reagent two factories under the chemical reagent head factory of Shanghai

KH ₂PO ₄

FW：136.09

10, potassium hydroxide (Potassium Hydroxide)

Shanghai development chemical reagent work

KOH

FW：56.10

11, magnesium sulfate (Magnesium Sulfate)

MgSO ₄

FW：120.38

12, dexamethasone

Shanghai the 9th pharmaceutical factory

1ml contains 5mg

Claims

1, a kind of method of preparing multi-organ preservation solution is prepared under the aseptic condition of strictness, and adopts aseptic filtration method sterilization, and the concrete steps of pressing 1000ml amount process for preparation are as follows:

20～30 ℃ of of In room temperature is under, adds successively in 1 liter container:the distilled water of 400ml; Add the 35.83g lactobionic acid, it is clear that stirring becomes until liquid color; Add 20ml 5N potassium hydroxide, and stirred 5 minutes; Add 5.0ml 5N sodium hydroxide, and stirred 5 minutes; Add adenosine 1.34g, it is clear stirring until liquid color; Add the 0.136g allopurinol, it is clear that stirring becomes until liquid color; Add the 3.4g potassium dihydrogen phosphate, it is clear that stirring becomes until liquid color; Add 0.60g magnesium sulfate, it is clear that stirring becomes until liquid color; Add 20.538g sucrose, it is clear that stirring becomes until liquid color; Add 0.92g glutathione reduced form, it is clear that stirring becomes until liquid color; The isoptin that adds 20mg, it is clear that stirring becomes until liquid color; Add the 8mg dexamethasone, it is clear that stirring becomes until liquid color; Add dextran-40 500ml (10%), heat and dissolve; As conditioning agent, pH is adjusted to 7.45 ± 0.10 with the sodium hydroxide of 5N; Add distilled water to 1000ml, and stirred 5 minutes; It is characterized in that its composition calculates content range by every liter and is: lactobionic acid 99-101mmol potassium dihydrogen phosphate 24-26mmol magnesium sulfate 4-6mmol adenosine 4-6mmol glutathione reduced form 2-4mmol dexamethasone 8mg allopurinol 0.5-1.5mmol5N NaOH 4-6ml5N potassium hydroxide 19-21ml sucrose 50-70mmol dextran-40 40-60g isoptin 20mg