CN1903177A - Alprostadil nano-particles and its prepn. method - Google Patents

Abstract

A nanoparticle of prostaglandin El contains prostaglandin El, the carrier chosen from poloxamer cholesterol ester of fatty acid and cholesterol ester of fatty acid, stabilizer and water. Its preparing process is also disclosed.

Description

Alprostadil nano-particles and preparation method thereof

Technical field

The present invention relates to a kind of Alprostadil nano-particles and preparation method thereof, it is characterized in that comprising Alprostadil, carrier, stabilizing agent and water, wherein carrier has been selected novel adjuvant fatty acid poloxamer cholesteryl ester (PPGC) and fatty acid PEG cholesteryl ester (PEGP) for use, nanoparticle particle diameter≤the 1um of preparation has the advantage of drug loading height and good stability.

Background technology

Alprostadil has another name called PGE1, and from 20 carbon fatty acids, endogenous PGE1 can be produced by nearly all cells in vivo.It is the broad spectrum activity material that exists in the human body, effect with intensive blood vessel dilating and anticoagulant, but to early stage vascular lesion microcirculation improvement, occlusive vascular disease is become the foundation that can impel collateral circulation, thereby allevating angina pectoris reduces the generation of myocardial infarction and dwindles infarct size.Because of its effect is rapid, curative effect is sure, is used for the treatment of coronary heart disease more and more.Sweden medical research personnel obtain the nineteen eighty-two Nobel Prize in medicine because of finding and study Alprostadil.

External advanced pharmaceutical technology, alprostadil injection (fat milk) are introduced in the pharmacy of Taide, nineteen ninety-five Beijing.From the clinical practice situation, alprostadil injection (fat milk) is compared with freeze-dried powder, solves the insufferable blood vessel irritation problem of freeze-dried powder, is subjected to clinical doctor and patient's favorable comment deeply.But because PGE1 is unstable under aqueous solution state, degraded easily so need stored refrigerated, Refrigerated Transport, so just makes the use cost of alprostadil injection not only increase but also very inconvenient, and effect duration one year.

Polyethylene Glycol (PEG) is the mixture that oxirane and water polycondensation form, and its molecular weight ranges is 1500-40000.

Poloxamer (Poluoshamu) trade name is pluronic (pluronic) F-68.Be white or the translucent waxy solid of little yellow, little have a foreign odor.In water and ethanol in easily molten, in dehydrated alcohol and ethyl acetate, dissolve.Purposes: as surfactant, emulsifying agent, suppository base, excipient etc.Poloxamer prevents to can be used for intravenous formulations by haemolysis because of having significantly, can play the hydrotropy effect to some medicine.

Poloxamer molecular formula: H (C2H4O) a (C3H6O) b (C2H4O) cOH, wherein a and c are 2～130, and b is 15～67, and containing polyoxyethylene is 81.8% ± 1.9%.Mean molecule quantity be 1000～7000 should be 90.0%～110.0% of labelled amount.Mean molecule quantity is 7000.

Cholesterol claims cholesterol again, is a kind of derivant of cyclopentanoperhy drophenanthrene, by staying body portion and a long side chain to form.The Chang Zuowei stabilizing agent uses in nanometer formulation.

Given this, of the present invention group of requirement according to preparation, adopted by poloxamer (POL) or Polyethylene Glycol (PEG) and cholesterol with fat diacid or the bonded novel adjuvant (Poloxamer-G-Cholesterol of fatty acid anhydride, POGC) or (PEG-G-Cholesterol, PEGC), and with prepared Alprostadil nano-particles as main adjuvant, because new dressing water-wet side poloxamer and the good emulsification property of PEG and hydrophobic section cholesterol and Alprostadil are with the good combination of molecular separating force, improve the medicine carrying ability of nanoparticle greatly, guaranteed the stable existence of medicine in nanoparticle.

The present invention adopts novel adjuvant that Alprostadil is made the solid nano grain, has overcome the shortcoming of using and store inconvenience (room temperature is preserved, 2 years effect duration) of alprostadil injection, and comparing with alprostadil injection more has degradation in vivo advantage slowly.

The present invention adopts high-pressure emulsification low-temperature setting method, is carrier material with POGC or PEGC, the preparation Alprostadil nano-particles, and investigate its physical stability, this work does not appear in the newspapers both at home and abroad at present.

Summary of the invention

Alprostadil nano-particles of the present invention and preparation method thereof, it comprises Alprostadil, carrier, stabilizing agent and water, and its parts by weight are:

Alprostadil 0.0005-0.05

Carrier 0.5-10

Stabilizing agent 1.0-20

Water 70-99

Wherein carrier is selected the cholesteryl ester that Biodegradable Polymers is modified, Biodegradable Polymers wherein, specifically refer to a kind of in poloxamer, the Polyethylene Glycol, the material of formation is called after fatty acid poloxamer cholesteryl ester (POGC) and fatty acid PEG cholesteryl ester (PEGC) respectively; Wherein the fatty acid carbons atomic number is 2-10.

Stabilizing agent select lecithin, triglyceride, mono fatty acid glyceride, in one or more; Specifically comprise bean bandit acid, Palmic acid, wych-elm acid, sad, capric acid, glyceryl monostearate, single trimyristin, monopalmitin, single Rikemal B 200, single caprylin, single caprin, glyceryl monostearate, two trimyristins, glycerol-1,3-dipalmitate, two Rikemal B 200s, two caprylins, two caprins, glycerol stearate, bean bandit acid triglyceride, Trihexanoylglycerol, wych-elm acid triglyceride, sad triglyceride, the capric acid triglyceride; Lecithin comprises soybean phospholipid, egg yolk lecithin, hydrogenated soya phosphatide, hydrogenation egg yolk lecithin, two nutmeg phosphatidylcholine, two nutmeg phosphatidyl glycerol, dipalmitoyl phosphatidyl choline, distearoyl phosphatidylcholine and distearyl phosphatidyl glycerol.

Alprostadil nano-particles of the present invention can also contain freeze drying protectant, is selected from glucose, sorbitol, lactose, maltose, mannitol, trehalose and the low molecular dextran one or more

Alprostadil nano-particles of the present invention, the wherein particle diameter≤1um of nanoparticle.

The present invention with novel adjuvant poloxamer (POL) or Polyethylene Glycol (PEG) and cholesterol with fat diacid or the bonded novel adjuvant (Poloxamer-G-Cholesterol of fatty acid anhydride, POGC) or (PEG-G-Cholesterol, PEGC) its synthetic method is as follows:

3～10g cholesterol and 0.1～1g 4-dimethylamino pyridine (DMAP) are placed 0.01～1ml dichloromethane (perhaps DMF), stir, slowly add 0.5～2.0g fatty acid anhydride, nitrogen protection is reaction 2h down, order adds 20-50g Polyethylene Glycol or 100～150g poloxamer and 1.0～3.0g dicyclohexylcarbodiimide (DCC) behind the natural cooling, slowly stirs 24hr to 1 week, rotary evaporation after reaction is finished, product dissolves with chloroform, gets crude product through ether sedimentation; Again through refining fatty acid poloxamer cholesteryl ester or the fatty acid polyglycol ethylene glycol cholesteryl ester of promptly getting of repeated precipitation.

Wherein the carbon number of fatty acid is 2-10.

The preparation technology of Alprostadil nano-particles:

Alprostadil, POGC or PEGP and the stabilizing agent heating of recipe quantity are dissolved, add in an amount of sterilized water for injection, stir and make dissolving; Mix above-mentioned two kinds of systems, add water to 100ml, high-speed stirred gets colostrum; Under the logical condition of nitrogen gas, the colostrum for preparing is met granularity requirements with the high pressure homogenizer homogenizing to detecting through laser particle analyzer, regulate pH value 5.0～7.0; the freeze drying protectant that adds recipe quantity in the aforesaid liquid, stirring and dissolving, aseptic filtration; fill, lyophilization are promptly.

Alprostadil nano-particles is the white loose block, and the back mean diameter of redissolving is controlled at below the 0.3 μ m, and the envelop rate of Alprostadil reaches more than 95%.

Specific embodiment

Embodiment one

Prescription: Alprostadil 0.0005-0.05g, POGC0.5～10.0g, glyceryl monostearate 1.0～6.0g, fatty acid 1.0～6.0g, lecithin 2.0～6.0g, mannitol 2.0～10.0g, water for injection adds to 30ml.

Preparation method one:

A) with glyceryl monostearate 2.0g, fatty acid 1.0g, lecithin 1.0g, POGC2.0g and Alprostadil 1mg heating and melting, colostrum is made in the water for injection (PH10-11) and the mannitol 5.0g stirring that add recipe quantity, regulate pH value 4.5～7.0, the reuse high pressure homogenizer is up to specification to granularity to the high pressure homogenize emulsifying that circulates of colostrum system.

B) after above-mentioned Emulsion filters, lyophilization, the inflated with nitrogen gland both got.

Preparation method two:

A) with glyceryl monostearate 3.0g, fatty acid 1.0g, lecithin 1.0g, POGC 10.0g and Alprostadil 50mg heating and melting, colostrum is made in the water for injection (PH10-11) and the mannitol 10.0g stirring that add recipe quantity, regulate pH value 4.5～7.0, the reuse high pressure homogenizer is up to specification to granularity to the high pressure homogenize emulsifying that circulates of colostrum system.

B) after above-mentioned Emulsion filters, lyophilization, the inflated with nitrogen gland both got.

Embodiment two

Prescription: Alprostadil 0.0005-0.05g, PEGC1.0～10.0g, glyceryl monostearate 1.0～6.0g, lecithin 2.0～6.0g, mannitol 2.0～6.0g, water for injection adds to 30ml.

A) with glyceryl monostearate 1.0g, lecithin 1.0g, PEGC3.0g and Alprostadil 0.0005g heating and melting, colostrum is made in the water for injection (PH10-11) and the mannitol 5.0g stirring that add recipe quantity, regulate pH value 4.5～7.0, the reuse high pressure homogenizer is up to specification to granularity to the high pressure homogenize emulsifying that circulates of colostrum system.

B) after above-mentioned Emulsion filters, lyophilization, the inflated with nitrogen gland both got.

Embodiment three

Prescription: Alprostadil 0.0005-0.05g, POGC 1.0～8.0g, mono laurate acid glyceride 1.0～6.0g, lecithin 1.0～3.0g, mannitol 2.0～6.0g, low molecular dextran 2.0～6.0g, water for injection adds to 100ml.

Alprostadil 0.005g, POGC0.5g, mono laurate acid glyceride 2.0g and lecithin 1.0g heating dissolve; add in the suitable quantity of water solution of mannitol 3.0g; stir; high-speed stirred is to emulsion droplet mean diameter≤0.5 micron; add freeze drying protectant low molecular dextran 2.0g; behind the dissolution filter, lyophilization, the inflated with nitrogen gland both got.

Embodiment four

Prescription: Alprostadil 0.0005-0.05g, PEGC 1.0～8.0g, single caprin 1.0～6.0g, mannitol 2.0～6.0g, sodium alginate 2.0～10.0g, water for injection adds to 30ml.

A) with Alprostadil 0.005g, the PEGC5.0g of recipe quantity, single caprin 2.0g heating and melting, colostrum is made in the water for injection and the mannitol 5.0g stirring that add recipe quantity, regulate pH value 4.5～7.0, the reuse high pressure homogenizer circulates high pressure homogenize emulsifying to granularity≤0.5 micron to the colostrum system; Add an amount of freeze drying protectant sodium alginate 5.0g, after the filtration, lyophilization, the inflated with nitrogen gland both got.

Embodiment five (blood vessel irritation test)

Trial drug: according to the preparation of embodiment (, two, four, five) provider's method, Alprostadil freeze-dried powder pin is buied from market, is mixed with 5% solution with 0.9% sodium chloride injection during test.

Experimental animal: healthy rabbits, body weight 2.0～2.2kg.

Test method: get 12 of healthy rabbits, male and female half and half.By body weight and sex grouping, 0.9% sodium chloride injection is matched group, alprostadil injection group and embodiment (one, two, three, a four) group, 2 every group, ear ear edge is pressed clinical administration concentration intravenous drip 10ml/kg in a rabbit left side, drip velocity 1ml/ branch, every day 1 time, continuous 7 days.Matched group is with method intravenous drip 0.9% sodium chloride injection.Observe the administration topical manifestations during except that each administration and after the administration, after the last intravenous drip, cut the medicine exterior feature of picking up the ears, conventional fixing after, go into pin proximal part 1cm place in the distance intravenous drip, cut the wide specimen of 0.5cm every 1cm, get 3 specimen altogether.Pathological observation under the mirror is carried out in section statining, the results are shown in following table:

The blood vessel irritation test

Project	The wide vasodilation of the rabbit ear	Red and swollen	Have or not cell infiltration
				0.9% sodium chloride injection matched group	-	-	-
Alprostadil freeze-dried powder pin	+++	+++	++
				Embodiment one	-	-	-
Embodiment two	-	-	-
				Embodiment three	-	-	-
Embodiment four	-	-	-

Remarks: " ++ " is serious, "+" a little, "-" do not have

Above result of the test shows that the preparation of the present invention's preparation has bland advantage.

Embodiment six (medicine for experiment relatively)

Trial drug:, be mixed with the solution that contains Alprostadil 5ug/ml with 0.9% sodium chloride injection during test according to the preparation of embodiment () provider method; Alprostadil injection is buied from market, specification: 5ug/ml.

Experimental animal: healthy rat, body weight 0.15～0.2kg, totally 20,10 every group.

Administration: 5ml/Kg

The blood sampling time point: 6,15,30,60min, 2,4,6,9h

Conclusion: nanoparticle of the present invention is compared with commercially available alprostadil injection, onset time basically identical, removing speed is slower in vivo, has played the effect of efficacy enhancing and toxicity reducing, the results are shown in Figure of description 1.

Description of drawings: Mean-F1 prepares according to embodiment () provider method for the present invention

Mean-F2 is alprostadil injection (commercially available)

Claims (7)

1, a kind of Alprostadil nano-particles and preparation method thereof is characterized in that comprising Alprostadil, carrier, stabilizing agent and water, and its parts by weight are:

Alprostadil 0.0005-0.05

Carrier 0.5-10

Stabilizing agent 1.0-20

Water 70-99.

2,, it is characterized in that the cholesteryl ester of selecting Biodegradable Polymers to modify according to the carrier in the Alprostadil nano-particles in the claim 1; Concrete select a kind of in poloxamer and the Polyethylene Glycol, the material of formation is respectively fatty acid poloxamer cholesteryl ester (POGC) and fatty acid polyglycol ethylene glycol cholesteryl ester (PEGC), and wherein the fatty acid carbons atomic number is 2-10.

3,, it is characterized in that selecting in lecithin, triglyceride, mono fatty acid glyceride and the difatty acid glyceride one or more according to the stabilizing agent in the Alprostadil nano-particles in the claim 1.

4,, it is characterized in that to comprise freeze drying protectant: one or more in glucose, sorbitol, lactose, maltose, mannitol, trehalose and the low molecular dextran according to the Alprostadil nano-particles in the claim 1.

5, according to the Alprostadil nano-particles in the claim 1, its preparation method is: Alprostadil, POGC or PEGC and the stabilizing agent heating of recipe quantity are dissolved, add an amount of sterilized water for injection, stir and make dissolving; Mix above-mentioned two kinds of systems, add water to 100ml, high-speed stirred gets colostrum; Under the logical condition of nitrogen gas, the colostrum for preparing is met granularity≤1um with high pressure homogenizer to detecting through laser particle analyzer, aseptic filtration, fill, lyophilization are promptly.

6, according to the fatty acid in the claim 3, it is characterized in that selecting satisfied fatty acid, specifically comprise bean bandit acid, Palmic acid, wych-elm acid, sad, capric acid, glyceryl monostearate, single trimyristin, monopalmitin, single Rikemal B 200, single caprylin, single caprin, glyceryl monostearate, two trimyristins, glycerol-1,3-dipalmitate, two Rikemal B 200s, two caprylins, two caprins, glycerol stearate, bean bandit acid triglyceride, Trihexanoylglycerol, wych-elm acid triglyceride, sad triglyceride, the capric acid triglyceride.

7,, it is characterized in that described lecithin comprises soybean phospholipid, egg yolk lecithin, hydrogenated soya phosphatide, hydrogenation egg yolk lecithin, two nutmeg phosphatidylcholine, two nutmeg phosphatidyl glycerol, dipalmitoyl phosphatidyl choline, distearoyl phosphatidylcholine and distearyl phosphatidyl glycerol according to the lecithin in the claim 3.

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