CN116649327A - Application of HMG-CoA and pharmaceutically acceptable salts thereof in preparation of platelet preservative - Google Patents
Application of HMG-CoA and pharmaceutically acceptable salts thereof in preparation of platelet preservative Download PDFInfo
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- CN116649327A CN116649327A CN202310499183.5A CN202310499183A CN116649327A CN 116649327 A CN116649327 A CN 116649327A CN 202310499183 A CN202310499183 A CN 202310499183A CN 116649327 A CN116649327 A CN 116649327A
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- CABVTRNMFUVUDM-VRHQGPGLSA-N (3S)-3-hydroxy-3-methylglutaryl-CoA Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)[C@@H](O)C(=O)NCCC(=O)NCCSC(=O)C[C@@](O)(CC(O)=O)C)O[C@H]1N1C2=NC=NC(N)=C2N=C1 CABVTRNMFUVUDM-VRHQGPGLSA-N 0.000 title claims abstract description 45
- 150000003839 salts Chemical class 0.000 title claims abstract description 20
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 239000003755 preservative agent Substances 0.000 title claims abstract description 6
- 230000002335 preservative effect Effects 0.000 title claims description 4
- WHBMMWSBFZVSSR-UHFFFAOYSA-N 3-hydroxybutyric acid Chemical compound CC(O)CC(O)=O WHBMMWSBFZVSSR-UHFFFAOYSA-N 0.000 claims description 9
- 239000002504 physiological saline solution Substances 0.000 claims description 7
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical class [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 claims description 3
- JFCQEDHGNNZCLN-UHFFFAOYSA-N glutaric acid Chemical compound OC(=O)CCCC(O)=O JFCQEDHGNNZCLN-UHFFFAOYSA-N 0.000 claims description 2
- 230000006907 apoptotic process Effects 0.000 abstract description 10
- 238000001802 infusion Methods 0.000 abstract description 10
- 238000001727 in vivo Methods 0.000 abstract description 9
- 230000004083 survival effect Effects 0.000 abstract description 8
- 230000002439 hemostatic effect Effects 0.000 abstract description 6
- 230000010118 platelet activation Effects 0.000 abstract description 3
- 210000001772 blood platelet Anatomy 0.000 description 93
- 238000004321 preservation Methods 0.000 description 20
- 230000000694 effects Effects 0.000 description 11
- 238000003860 storage Methods 0.000 description 11
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 9
- 108090000672 Annexin A5 Proteins 0.000 description 8
- 102000004121 Annexin A5 Human genes 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 8
- 238000000338 in vitro Methods 0.000 description 8
- 241000699670 Mus sp. Species 0.000 description 7
- 238000011579 SCID mouse model Methods 0.000 description 7
- 101000622137 Homo sapiens P-selectin Proteins 0.000 description 6
- 102100023472 P-selectin Human genes 0.000 description 6
- 238000007619 statistical method Methods 0.000 description 6
- 239000000243 solution Substances 0.000 description 5
- 206010007688 Carotid artery thrombosis Diseases 0.000 description 4
- 208000007536 Thrombosis Diseases 0.000 description 4
- 230000004913 activation Effects 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- -1 tri-hydroxy trimethyl glutaryl Chemical group 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- 108010040476 FITC-annexin A5 Proteins 0.000 description 2
- 101001078143 Homo sapiens Integrin alpha-IIb Proteins 0.000 description 2
- 101001070790 Homo sapiens Platelet glycoprotein Ib alpha chain Proteins 0.000 description 2
- 102100025306 Integrin alpha-IIb Human genes 0.000 description 2
- 102100034173 Platelet glycoprotein Ib alpha chain Human genes 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000001715 carotid artery Anatomy 0.000 description 2
- VYXSBFYARXAAKO-WTKGSRSZSA-N chembl402140 Chemical compound Cl.C1=2C=C(C)C(NCC)=CC=2OC2=C\C(=N/CC)C(C)=CC2=C1C1=CC=CC=C1C(=O)OCC VYXSBFYARXAAKO-WTKGSRSZSA-N 0.000 description 2
- RGJOEKWQDUBAIZ-UHFFFAOYSA-N coenzime A Natural products OC1C(OP(O)(O)=O)C(COP(O)(=O)OP(O)(=O)OCC(C)(C)C(O)C(=O)NCCC(=O)NCCS)OC1N1C2=NC=NC(N)=C2N=C1 RGJOEKWQDUBAIZ-UHFFFAOYSA-N 0.000 description 2
- 239000005516 coenzyme A Substances 0.000 description 2
- 229940093530 coenzyme a Drugs 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- KDTSHFARGAKYJN-UHFFFAOYSA-N dephosphocoenzyme A Natural products OC1C(O)C(COP(O)(=O)OP(O)(=O)OCC(C)(C)C(O)C(=O)NCCC(=O)NCCS)OC1N1C2=NC=NC(N)=C2N=C1 KDTSHFARGAKYJN-UHFFFAOYSA-N 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000005976 HMG-CoA lyase Human genes 0.000 description 1
- 108020003145 HMG-CoA lyase Proteins 0.000 description 1
- QGMRQYFBGABWDR-UHFFFAOYSA-M Pentobarbital sodium Chemical compound [Na+].CCCC(C)C1(CC)C(=O)NC(=O)[N-]C1=O QGMRQYFBGABWDR-UHFFFAOYSA-M 0.000 description 1
- 241000220156 Saxifraga Species 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 238000002617 apheresis Methods 0.000 description 1
- 230000020411 cell activation Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 210000003593 megakaryocyte Anatomy 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229960002275 pentobarbital sodium Drugs 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 210000003670 sublingual gland Anatomy 0.000 description 1
- 230000001732 thrombotic effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
-
- 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
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/19—Platelets; Megacaryocytes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0644—Platelets; Megakaryocytes
Abstract
The invention discloses application of HMG-CoA and pharmaceutically acceptable salts thereof in preparation of platelet preserving agents. The invention proves that the addition of HMG-CoA or the pharmaceutically acceptable salt thereof to the platelet to be preserved can effectively relieve platelet activation and apoptosis, prolong the in vivo survival period of the platelet after platelet infusion and protect the hemostatic function of the platelet.
Description
Technical Field
The invention relates to a platelet preservation technology, in particular to application of tri-hydroxy trimethyl glutaryl coenzyme A and pharmaceutically acceptable salts thereof in preparation of platelet preservation agent.
Background
Platelets are small masses of cytoplasm formed by degranulation of megakaryocytes, and have a major function of participating in anticoagulation. Although there are several reasons for platelet supply shortage, the too short shelf life (only 5 days) under conventional storage conditions (22±2 ℃ continuous shaking) is one of the major factors, which directly results in an outdated rejection rate of up to 10 to 20%. Because of the high outdated rejection rate, the shortage of the special equipment and the high price of consumable materials required by platelet collection are considered, and a great amount of blood preparation of platelets is difficult to develop in each blood center and each blood station, and the shortage is further aggravated by the high outdated rejection rate.
Disclosure of Invention
In view of the shortcomings or drawbacks of the prior art, the present invention provides the use of HMG-CoA and its pharmaceutically acceptable salts for the preparation of platelet preserving agents. The invention also provides corresponding platelets, and HMG-CoA or pharmaceutically acceptable salts thereof are additionally arranged in the platelets.
Alternatively, the pharmaceutically acceptable salt of HMG-CoA is the disodium salt of tri-hydroxy trimethyl glutarate monoacyl-CoA. The addition of HMG-CoA or a pharmaceutically acceptable salt thereof to platelets does not result in a beta-hydroxybutyric acid content in platelets exceeding 0.27mmol/L. Preferably, the concentration of HMG-CoA or a pharmaceutically acceptable salt thereof in platelets is 0.2mmol/L or less.
The invention proves that the addition of HMG-CoA or the pharmaceutically acceptable salt thereof to the platelet to be preserved can effectively relieve platelet activation and apoptosis, prolong the in vivo survival period of the platelet after platelet infusion and protect the hemostatic function of the platelet.
Drawings
FIG. 1 shows the effect of HMG-CoA.2Na on PS eversion during in vitro platelet Preservation (PS) in example 1 of the present invention + Platelet percentage statistics); n=6, all data in the histogram using mean±sem; NS: no statistical difference; * P<0.01。
FIG. 2 is a graph showing the effect of HMG-CoA.2Na on CD62P expression during in vitro platelet preservation (CD 62P) in example 2 of the present invention + Platelet percentage statistics); n=6, all data in the histogram using mean±sem; NS: no statistical difference; * P<0.001。
FIG. 3 shows the effect of HMG-CoA.2Na on in vivo survival after platelet infusion (APC) in example 3 of the present invention + /APC + &FITC + Platelet analysis statistics); n=5, all data in the histogram using mean±sem; NS: no statistical difference; * P is p<0.05；**p<0.01。
FIG. 4 is a graph showing the effect of HMG-CoA.2Na on in vivo hemostatic function after platelet infusion (mouse carotid thrombosis time statistic) in example 4 of the present invention; n=6, ns: no statistical difference; * p <0.05.
Detailed Description
Unless specifically stated otherwise, scientific and technical terms herein have been understood based on the knowledge of one of ordinary skill in the relevant art.
It is well known that platelet preservation damage (platelet storage lesions, PSLs) occurs during storage of platelets, and is manifested by apoptosis and activation of cells with prolonged storage time, shortening of in vivo survival after platelet preservation infusion, and impaired anticoagulant function. Therefore, firstly, the influence of HMG-CoA on the PS eversion of human platelets is analyzed, and the HMG-CoA has the effect of relieving the apoptosis in the platelet in-vitro preservation process; the effect of HMG-CoA on human platelet CD62P expression was then analyzed, demonstrating that HMG-CoA has the effect of alleviating activation during in vitro platelet preservation. Based on this, the inventors further analyzed the ratio of HMG-CoA pretreatment human platelets infused into SCID mice and carotid thrombosis time of SCID mice infused with HMG-CoA pretreatment human platelets by a SCID mouse platelet infusion model, and verified that HMG-CoA can increase in vivo survival after platelet infusion while protecting hemostatic function.
The invention is further illustrated by the following examples. It should be noted that, tri-hydroxy trimethyl glutaryl CoA (HMG-CoA) is one of the metabolic intermediates of human fat oxidation, and can be converted into beta-hydroxybutyric acid after the action of HMG-CoA lyase and beta-hydroxybutyrate dehydrogenase, respectively. In practice, HMG-CoA is usually present in the form of a salt, and the hydrolysis product of HMG-CoA salt is HMG-CoA, and the following examples are presented by taking tri-hydroxy trimethyl glutaryl coenzyme A disodium salt as a representative for illustrating the present invention. In addition, platelets used in the examples below were all obtained from healthy, gratuitous donors, HMG-CoA.2Na from Sigma, annexin V-FITC Apoptosis Staining/Detection from BD; male SCID mice were purchased from Saxifraga, 20-25g, and both APC anti-human CD41 anti-body and FITC anti-mouse CD41 anti-body were purchased from Biolegend; anti-mico CD42b Anti body was purchased from Emfret and rhodamine was purchased from Sigma.
Example 1 the inventors have found that HMG-CoA.2Na has the effect of alleviating apoptosis during in vitro storage of human platelets
1. Preparation of platelets
Healthy adults who did not take any medication for 10 days were recruited as volunteers and apheresis platelets were prepared for later use using a hemocytometer (Amicus 4r4580, fenwal).
2. Platelet processing
Three platelet-preserving bags were added by sucking 5mL of platelets respectively, one of which was directly used as a control group 1, the other was added with 25. Mu.L of physiological saline (vehicle) as a control group 2, and the third was added with 25. Mu.L of physiological saline (final concentration of HMG-CoA.2Na in platelet-processing system is 0.2 mmol/L) dissolved with HMG-CoA.2Na as an experimental group; three groups of platelets are placed outside a platelet shake preservation box for preservation (continuous shake at 22+/-2 ℃).
PS-Positive platelet detection
Appropriate amounts of platelets were taken at day 3, 5, and 7 of storage, respectively, and PS were labeled with reference to Annexin V-FITC Apoptosis Staining/Detection instructions, and then the Annexin V-positive platelet percentage was detected using a flow cytometer (BD FACS Canto).
As shown in the figure 1, the statistical analysis of the percentage of the Annexin V-positive blood platelets is that the percentage of the Annexin V-positive blood platelets in the control groups 1 on days 3, 5 and 7 is 6.2%, 8.5% and 10.6%, respectively, and the statistical analysis of the percentage of the Annexin V-positive blood platelets in the control groups 2 on days 3, 5 and 7 is not different from the statistical analysis of the percentage of the Annexin V-positive blood platelets in the control groups on days 6.2%, 8.4% and 10.7% respectively; the results demonstrate that the addition of an amount of physiological saline does not affect the percentage of Annexin V positive platelets.
The percentages of Annexin V positive platelets on days 3, 5 and 7 of the experimental group were 4.2%, 5.3%, 8.0%, respectively, all significantly lower than 6.2%, 8.5%, 10.6% on days 3, 5 and 7 of the control group 1, and 8.0% on day 7 of the experimental group was not statistically different from 8.5% on day 5 of the control group 1; the results demonstrate that the addition of an amount of HMG-CoA.2Na reduced the percentage of Anexin V-positive platelets on day 7 of the experimental group, which was similar to that on day 5 of the control group 1.
Annexin V is a fluorescent probe for PS, and PS eversion is a classical indicator of apoptosis. Thus, the above results of this example demonstrate that HMG-coa.2na has the effect of alleviating apoptosis of human platelets during in vitro storage, potentially extending the shelf life of platelets from the conventional 5 days to 7 days.
And the continuous monitoring method of enzyme dynamics is adopted for detection, when HMG-CoA.2Na with the final concentration of 0.2mmol/L is added into the platelets in the experimental group of the embodiment, the beta-hydroxybutyric acid in the platelet preservation system is raised to 0.22mmol/L and not to exceed 0.27mmol/L of the upper limit of the normal value of the human body. Therefore, the invention can preserve the platelet by adding a certain amount of HMG-CoA.2Na, and does not bring harm to the organism.
Example 2 the inventors have found that HMG-CoA.2Na has an effect of alleviating activation during in vitro storage of human platelets
Preparation and treatment of platelets the platelets were prepared and treated as in example 1, and appropriate amounts of platelets were taken on days 3, 5 and 7, respectively, and the percentage of CD62P positive platelets was detected using a flow cytometer (BD FACS Canto) after FITC anti-human CD62P labelling.
As shown in FIG. 2, the statistical analysis of the percentage of CD 62P-positive platelets is that the percentage of CD 62P-positive platelets is 11.8%, 15.8%, 25.9% on days 3, 5 and 7 of the control group 1, and no statistical difference is found from the percentage of CD 62P-positive platelets 11.9%, 15.9%, 25.8% on days 3, 5 and 7 of the control group 2, which indicates that the addition of a certain amount of physiological saline does not affect the percentage of CD 62P-positive platelets.
The percentages of CD62P positive platelets on days 3, 5 and 7 of the experimental group were 8.6%, 10.3%, 16.9%, respectively, all significantly lower than 11.8%, 15.8%, 25.9% on days 3, 5 and 7 of the control group 1, and 16.9% on day 7 of the experimental group was not statistically different from 15.8% on day 5 of the control group 1. The results demonstrate that the addition of an amount of HMG-CoA.2Na reduced the percentage of CD 62P-positive platelets, similar to that of control group 1, day 5, on day 7 of the experimental group.
CD62P production is a classical indicator of platelet activation, and thus, the above results of this example demonstrate that HMG-coa.2na has the effect of delaying activation of human platelets during in vitro storage, potentially extending shelf life of platelets.
EXAMPLE 3 the inventors have found that HMG-CoA.2Na pretreatment can prolong the in vivo survival after platelet transfusion
Example 1 and example 2 above demonstrate that the addition of an amount of HMG-coa.2na affects apoptosis and activation of platelets, while physiological saline does not, and thus this example reduces control group 2 on the basis of example 1, and platelets are prepared and treated as in example 1;
then, on the 5 th day of preservation, the control group 1 platelets were taken 4×10 8 And 4X 10 platelets from the experimental group at day 5 and day 7 of storage, respectively 8 SCID mice were then transfused via the tail vein, 50. Mu.l of peripheral blood was obtained by cutting the tail tip at 0.5, 2, and 5 hours, and APCs were detected by flow cytometry (BD FACS Canto) using APC anti-human CD41Antibody and FITC anti-mouse CD41Antibody markers after erythrocyte lysis + /APC + &FITC + Percent.
Detected APC + /APC + &FITC + Percent statistical analysis As shown in FIG. 3, the APCs at 0.5, 2 and 5 hours after platelet storage of the experimental group on day 5 was infused into mice + /APC + &FITC + 41.2%, 32.0% and 25.4% respectively, which are significantly higher than the 26.0%, 18.9% and 17.5% of platelets stored in control group 1 at day 5, at hours 0.5, 2 and 5 after being introduced into mice;
and the experimental group stored the APCs at day 7, hours 0.5, 2 and 5 + /APC + &FITC + 24.5%, 19.2%, 16.3% of (C) were not statistically different from the control 1 at 26.0%, 18.9%, 17.5% of (C) 5.
Wherein APC + /APC + &FITC + The percentage represents the ratio of human platelets infused into SCID mice, which is proportional to the in vivo survival following platelet infusion, thus the results of this example demonstrate that HMG-coa.2na can prolong the in vivo survival following platelet infusion; it was also demonstrated that HMG-CoA.2Na pretreatment can extend the shelf life of platelets.
EXAMPLE 4 the inventors have found that HMG-CoA.2Na pretreatment protects the hemostatic function of platelets
Scid mouse platelet knockout
SCID mice were prepared, and autologous platelets were previously removed from the mice by injecting Anti-mice CD42b Antibody monoclonal Antibody via tail vein in the literature.
2. Platelet infusion
Preparation and treatment of platelets in this example were the same as in example 1, except that control group 2 was reduced; then, the control group 1 platelet was taken at the 5 th day of preservation, and the experimental group platelet was taken at the 5 th and 7 th days of preservation, respectively, each 1X 10 9 The mice were infused with the corresponding drug via the tail vein.
3. Mouse Fecl 3 Establishment of carotid thrombosis model
(1) Preparing 0.5mg/mL rhodamine 6G solution, and storing in a dark condition; (2) preparation of FeCl 100mg/mL 3 The solution is ready for use; (3) after the mice treated in the step 2 are anesthetized with 10G/L pentobarbital sodium solution, 200 mu L of rhodamine 6G solution is injected from the inner canthus of the mice; (4) the mouse is in a supine position, the skin of the mouse is cut along the median line of the neck, the sublingual gland is separated under a stereoscopic microscope, the mouse is turned up, and the carotid artery of the mouse is fully stripped and exposed; (5) 100mg/mL FeCl is dipped by a filter paper sheet with the length and the width of 1mm 3 The solution is stuck to the carotid artery of the mouse for 1min; (6) the filter paper sheet was removed, washed with physiological saline, and the thrombotic process was observed under a whole-body fluorescence microscopeThe thrombus formation time was recorded.
Statistical analysis of carotid thrombosis time in mice as shown in fig. 4, the average thrombosis time on day 5 of the experimental group was 490 seconds, significantly shorter than 898 seconds on day 5 of the control group 1, and there was no statistical difference between the average thrombosis time 905 seconds on day 7 of the experimental group and 898 seconds on day 5 of the control group 1. The thrombus formation time is inversely proportional to the platelet function, so the results of this example demonstrate that HMG-coa.2na can protect the hemostatic function of platelets; it was also further demonstrated that HMG-CoA.2Na pretreatment can extend the shelf life of platelets.
Claims (8)
- Use of hmg-CoA and its pharmaceutically acceptable salts for the preparation of a platelet preservative.
- 2. The use according to claim 1, wherein the HMG-CoA pharmaceutically acceptable salt is the disodium salt of tri-hydroxy trimethyl glutarate monoacyl-CoA.
- 3. The use according to claim 1, wherein the HMG-CoA or a pharmaceutically acceptable salt thereof is added to the platelets in an amount which does not result in a β -hydroxybutyrate content of more than 0.27mmol/L in the platelets.
- 4. The use according to claim 1, wherein the concentration of HMG-CoA or a pharmaceutically acceptable salt thereof in platelets is equal to or less than 0.2mmol/L.
- 5. A platelet preservative characterized by being prepared by dissolving HMG-CoA or a pharmaceutically acceptable salt thereof in physiological saline.
- 6. A platelet characterized in that HMG-CoA or a pharmaceutically acceptable salt thereof is additionally added to the platelet.
- 7. The platelet according to claim 6, wherein the HMG-CoA or a pharmaceutically acceptable salt thereof is added to the platelet in an amount which does not result in a β -hydroxybutyrate content in the platelet exceeding 0.27mmol/L.
- 8. The platelet according to claim 6, wherein the concentration of HMG-CoA or a pharmaceutically acceptable salt thereof in the platelet is 0.2mmol/L or less.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310499183.5A CN116649327A (en) | 2023-05-05 | 2023-05-05 | Application of HMG-CoA and pharmaceutically acceptable salts thereof in preparation of platelet preservative |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310499183.5A CN116649327A (en) | 2023-05-05 | 2023-05-05 | Application of HMG-CoA and pharmaceutically acceptable salts thereof in preparation of platelet preservative |
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| Publication Number | Publication Date |
|---|---|
| CN116649327A true CN116649327A (en) | 2023-08-29 |
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|---|---|---|---|
| CN202310499183.5A Pending CN116649327A (en) | 2023-05-05 | 2023-05-05 | Application of HMG-CoA and pharmaceutically acceptable salts thereof in preparation of platelet preservative |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116649327A (en) |
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- 2023-05-05 CN CN202310499183.5A patent/CN116649327A/en active Pending
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