CN105949278B - Substituted tetrahydrofuran water-soluble derivative and application thereof - Google Patents

Abstract

Description

Substituted tetrahydrofuran water-soluble derivative and application thereof

Technical Field

The invention belongs to the technical field of medicinal chemistry, and relates to a tetrahydrofuran derivative, in particular to a substituted tetrahydrofuran water-soluble derivative or a pharmaceutically acceptable acid salt, solvate or hydrate thereof, a preparation method thereof, a pharmaceutical composition thereof and application thereof in preparing a medicament for preventing or treating fungal infection.

Background

In application No. 5/30 in 1996, application No. 96190848.3 discloses a tetrahydrofuran fungicide, posaconazole (posaconazole) has been approved by the FDA in the united states for use in refractory diseases or other drug-resistant fungal infections (e.g., aspergillosis, tuberculosis, fusarium, etc.), and posaconazole oral suspensions have been approved by the food and drug administration in china. However, due to the structural characteristics of the compounds, the water solubility is poor, the intravenous administration preparation needs to be prepared into a test solution with the pH value of 2.6, the peripheral intravenous administration has obvious vascular irritation, the central intravenous administration is needed in clinical use to reduce the occurrence of phlebitis, and inconvenience and pain are brought to patients.

There is a need for broad spectrum antifungal agents with enhanced solubility and greater activity for treating systemic fungal infections, particularly Aspergillus, Candida, Cryptococcus and opportunistic infections, while having advantageous physical forms to produce formulations with more convenient and safe routes of administration.

Disclosure of Invention

The purpose of the invention is as follows: the invention aims to overcome the defects of the prior art and provides a substituted tetrahydrofuran water-soluble derivative for preparing a medicament for preventing or treating fungal infectious diseases; the derivatives of the substituted tetrahydrofuran have the characteristics of good water solubility, rapid conversion into active ingredients in vivo and suitability for conventional injections for peripheral intravenous administration.

The second object of the present invention is to provide a process for the preparation of the above-mentioned water-soluble derivatives of substituted tetrahydrofurans.

It is a third object of the present invention to provide a pharmaceutical composition comprising the above-mentioned water-soluble derivative of substituted tetrahydrofuran.

A fourth object of the present invention is to provide the use of the above-mentioned water-soluble derivative of substituted tetrahydrofuran or a pharmaceutical composition for the manufacture of a medicament for the prevention or treatment of fungal infections in a host, including warm-blooded animals, especially humans.

The technical scheme is as follows: the invention provides a substituted tetrahydrofuran water-soluble derivative, such as a compound shown in a general formula I, or a pharmaceutically acceptable acid salt thereof:

Wherein:

X1 is fluoro or chloro;

X2 is fluoro or chloro;

R1 is a dipeptide acyl or polypeptide acyl group wherein at least one aminoacyl group is formed from a natural amino acid; the compound is selected from one of the following compounds, or pharmaceutically acceptable acid salt thereof:

The derivatives provided by the invention comprise optical isomers of the compounds in the formula I.

The derivatives of the present invention include compounds of formula I or pharmaceutically acceptable acid salts thereof, including but not limited to the salts of the compounds with the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, citric acid, tartaric acid, phosphoric acid, lactic acid, acetic acid, maleic acid, fumaric acid, malic acid, mandelic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, oxalic acid or succinic acid.

The "amino acid acyl group" in the present invention refers to a group (NH2-R-C (═ O) -) formed by the amino acid (NH2-R-C (═ O) -OH) in which the carboxyl group is deficient in — OH. The amino acids referred to herein include 20 natural amino acids, for example: alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), proline (Pro), phenylalanine (Phe), tryptophan (Trp), methionine (Met), glycine (Gly), serine (Ser), threonine (Thr), cysteine (Cys), tyrosine (Tyr), asparagine (Asn), glutamine (gin), aspartic acid (Asp), glutamic acid (Glu), lysine (Lys), arginine (Arg), histidine (His) and the like, and also include various unnatural amino acids.

In a second aspect, the present invention provides a process for the preparation of a compound of formula I, or a pharmaceutically acceptable acid salt thereof, as a water-soluble derivative of the above-mentioned substituted tetrahydrofuran, comprising the steps of:

Referring to the synthesis method of patent CN 96190848.3, the compound of formula II was synthesized first using different starting materials A, B, C, D.

Reacting the compound of formula II with the compound of formula III in the presence of a condensing agent or reacting the compound of formula II with the compound of formula IV, and then removing the amino protecting group to obtain the compound of formula I

Wherein, X1 and X2 in the compound of formula II are defined as the compound of formula I, R3 is benzyloxycarbonyl protecting amino terminal, and the corresponding protecting group of the side chain with the corresponding protecting group on the amino acid side chain can be trityl, tert-butyloxycarbonyl, benzyloxycarbonyl, fluorenylmethyloxycarbonyl, etc., when the compound of formula III is used as the synthesis raw material in the preparation reaction, the condensing agent can be Carbonyldiimidazole (CDI), N, N ' -Diisopropylcarbodiimide (DIC), N, N ' -Dicyclohexylcarbodiimide (DCC), N- (3-dimethylaminopropyl) -N ' -ethylcarbodiimide hydrochloride (EDC. HCl), O- (7-azabenzotriazole) -N, N, N ', N ' -tetramethyluronium Hexafluorophosphate (HATU), benzotriazol-N, N, N ', N ' -tetramethyluronium Hexafluorophosphate (HBTU), corresponding catalysts such as N, N-dimethylamino pyridine and acid-binding agents can be added in the reaction, and a Cbz group is a benzyloxycarbonyl protecting group for protecting amino.

As a preferred embodiment, the present invention provides a process for the preparation of a compound of formula I or a pharmaceutically acceptable salt thereof, which comprises dissolving a compound of formula II or a salt thereof in an organic solvent, adding a base in portions while cooling, then reacting with a compound of formula IV, and removing the Cbz protecting group by catalytic hydrogenation to give a compound of formula I; or reacting the compound of formula III with a condensing agent, then with an organic solvent for the compound of formula II or a salt thereof, adding a base, and then removing the Cbz protecting group by catalytic hydrogenation to give the compound of formula I, which can be further purified by conventional methods such as recrystallization, column chromatography, etc., if necessary. Here, the base may be an inorganic base or an organic base, and may be selected from sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, triethylamine, or N, N-diisopropylethylamine, etc. Salifying the compound of the formula I with an organic solvent solution or an aqueous solution of an acid according to a ratio to obtain a salt of the compound of the formula I.

In particular, for compounds of the present invention in which the amino acid side chain has additional reactive functional groups, the side chain protected amino acid is used as the starting material, and a deprotection step is added after the condensation reaction.

In a third aspect, the present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I, or a pharmaceutically acceptable acid salt thereof, which may further comprise a pharmaceutically acceptable carrier or diluent. The pharmaceutical composition can also be combined with other active ingredients to form a combined composition or a composition with synergistic effect. The pharmaceutical composition can be administered by intravenous injection, by injection into tissue, intraperitoneal administration, oral administration or intranasal administration. The pharmaceutical composition may have a form selected from the group consisting of a solution, a dispersion, a suspension, a powder, a capsule, a tablet, a pill, an extended release capsule, an extended release tablet, and an extended release pill. The administration dosage of the pharmaceutical composition is 5-5000 mg/day.

In a fourth aspect, the present invention provides the use of a water-soluble derivative of a substituted tetrahydrofuran as defined above, such as a compound of formula I, or a pharmaceutically-acceptable acid salt thereof, in the manufacture of a medicament for the prophylaxis or treatment of a fungal infection in a host, including a warm-blooded animal, especially a human; wherein the fungus is selected from Candida, Histoplasma capsulatum, Sedum cerevisia, Zygomycetes bipolaris, Fusarium, Saccharomyces, and NonB. albicans strain, Cryptococcus neoformans or Aspergillus with fluconazole resistance.

Has the advantages that: compared with the prior art, the novel substituted tetrahydrofuran derivative has the characteristics of obvious antifungal infection activity, high bioavailability, very good water solubility (easy injection administration), convenient preparation, small irritation to blood vessels, no phlebitis caused by intravenous administration and the like.

Detailed Description

The technical solution of the present invention is described in detail by the following specific examples, and it is obvious to those skilled in the art that the embodiments of the present invention can be modified based on the prior art under the teaching of the present invention, and still fall into the protection scope of the present invention.

The compound starting sources used in the examples were: all reagents were purchased from a reagent company, the compound of formula II was synthesized using the method referenced CN 96190848.3 using different starting materials A, B, C, D, corresponding to II1, II2, II3, II4, respectively, posaconazole (posaconazole, II3) was purchased from a reagent company.

NMR data were collected and processed by a Bruker AV-300 NMR spectrometer.

example 1: synthesis of MJ10821

(1) Synthesis of intermediate M21

Cbz-Val-Ala-OH (322mg,1mmol) is dissolved in 3ml of N, N-Dimethylformamide (DMF), the temperature is reduced to-5 ℃, DIC (63.1mg,0.5mmol) is added under stirring, the reaction is carried out for 30min at room temperature, the temperature is further reduced to-5 ℃, 2ml of DMF solution of II3(350mg,0.5mmol), triethylamine (60.7mg,0.6mmol) and a catalytic amount of 4-Dimethylaminopyridine (DMAP) are sequentially added, the reaction is carried out for 4h at room temperature, after the reaction is finished, 15ml of water is poured, 3ml of ethyl acetate is used for extraction, organic layers are combined, anhydrous sodium sulfate is dried, a solvent is dried in a rotary manner, yellow solid is obtained, and the separation is carried out by silica gel column chromatography, so that 281mg of white-like powder (M21) is obtained, and. MS (m/z): 1005.5[ M +1] +.

(2) Synthesis of MJ10821

m21(251mg,0.25mmol) was dissolved in 3ml of methanol, 5% Pd/C200 mg was added, and the reaction was carried out under hydrogen gas for 30min, after completion of the reaction, Pd/C was filtered off, the solvent was spun off from the filtrate, and the filtrate was separated by silica gel column chromatography to obtain 128mg of white powder (MJ10821), with a yield of 58.8%. MS (m/z): 871.4[ M +1] +; 1H-NMR (DMSO-d6) delta: 0.76-0.81(d,6H), 0.85-0.88(t,3H),1.11-1.15(d,3H),1.17-1.27(d,3H),1.72-1.78(m,2H),1.93-2.02(m,1H),2.15-2.50(m,2H),2.51-2.61(m,1H),2.91-3.01(m,1H),3.09-3.41(m,8H),3.51-4.09(m,5H),4.10-4.29(m,2H),4.49-4.71(m,2H),5.01-5.21(m,2H),6.77-6.88(m,2H),6.89-6.97(m,2H), 6.98-7.7 (m,1H), 7.7-7.18H), 7.7-1.78 (m, 7H), 7.58-1.7 (m, 7.58-4.7H), 1H) 8.05-8.18(m,1H),8.28-8.42(m, 2H).

Example 2: synthesis of MJ10807

Reference is made to the synthesis of MJ10821, except that Cbz-Gly-Pro-OH and II3 are used as starting materials, which are condensed and deprotected to finally obtain MJ 10807. MS (m/z): 855.4[ M +1] +.

Example 3: synthesis of MJ10808

Reference is made to the synthesis of MJ10821, with the difference that starting materials Cbz-Ala-Pro-OH and II3 were selected, which were condensed and deprotected to finally obtain MJ 10808. MS (m/z): 869.4[ M +1] +.

Example 4: synthesis of MJ10809

Reference is made to the synthesis of MJ10821, which differs in that Cbz-Val-Pro-OH and II3 are selected as starting materials, and condensation and deprotection are carried out to finally obtain MJ 10809. MS (m/z): 897.5[ M +1] +.

Example 5: synthesis of MJ10819

Reference is made to the synthesis of MJ10821, except that Cbz-Gly-Ala-OH and II3 are used as starting materials, which are condensed and deprotected to obtain MJ 10819. MS (m/z): 829.4[ M +1] +.

Example 6: synthesis of MJ10820

Reference is made to the synthesis of MJ10821, except that Cbz-Ala-Ala-OH and II3 were used as starting materials, which were condensed and deprotected to give MJ 10820. MS (m/z): 843.4[ M +1] +.

Example 7: synthesis of MJ10825

Reference is made to the synthesis of MJ10821, except that Cbz-Leu-Ala-OH and II3 are used as starting materials, which are condensed and deprotected to give MJ 10825. MS (m/z): 885.5[ M +1] +.

Example 8: synthesis of MJ10826

Reference is made to the synthesis of MJ10821, except that Cbz-Ile-Ala-OH and II3 are used as starting materials, which are condensed and deprotected to finally obtain MJ 10826. MS (m/z): 885.5[ M +1] +.

example 9: synthesis of MJ10827

Reference MJ10821 was synthesized, except that Cbz-Phe-Ala-OH and II3 were used as starting materials, which were condensed and deprotected to finally obtain MJ 10827. MS (m/z): 919.4[ M +1] +.

Example 10: synthesis of MJ10830

Reference is made to the synthesis of MJ10821, except that Cbz-Asn-Ala-OH and II3 are used as starting materials, which are subjected to condensation and deprotection to finally obtain MJ 10830. MS (m/z): 886.4[ M +1] +.

Example 11: synthesis of MJ10833

Reference is made to the synthesis of MJ10821, except that Cbz-Gln-Ala-OH and II3 are used as starting materials, which are condensed and deprotected to give MJ 10833. MS (m/z): 900.4[ M +1] +.

Example 12: synthesis of MJ10839

Reference is made to the synthesis of MJ10821, except that Cbz-Pro-Ala-OH and II3 were used as starting materials, which were condensed and deprotected to give MJ 10839. MS (m/z): 869.4[ M +1] +.

Example 13: synthesis of MJ10844

Reference is made to the synthesis of MJ10821, except that Cbz-Ala-Gly-OH and II3 are used as starting materials, which are condensed and deprotected to obtain MJ 10844. MS (m/z): 829.4[ M +1] +.

Example 14: synthesis of MJ10845

Reference is made to the synthesis of MJ10821, which is different in that Cbz-Val-Gly-OH and II3 are selected as starting materials, and are subjected to condensation and deprotection to finally obtain MJ 10845. MS (m/z): 857.4[ M +1] +.

Example 15: synthesis of MJ10801

Reference is made to the synthesis of MJ10821, which differs in that Cbz-Gly-Pro-OH and II1 are selected as starting materials, and are subjected to condensation and deprotection to finally obtain MJ 10801. MS (m/z): 887.3[ M +1] +.

Example 16: synthesis of MJ10802

Reference is made to the synthesis of MJ10821, with the difference that Cbz-Ala-Pro-OH and II1 are used as starting materials, which are condensed and deprotected to finally obtain MJ 10802. MS (m/z): 901.4[ M +1] +.

Example 17: synthesis of MJ10803

reference is made to the synthesis of MJ10821, which differs in that Cbz-Val-Pro-OH and II1 are selected as starting materials, and are subjected to condensation and deprotection to finally obtain MJ 10803. MS (m/z): 929.4[ M +1] +.

Example 18: synthesis of MJ10813

Reference is made to the synthesis of MJ10821, except that Cbz-Gly-Ala-OH and II1 are used as starting materials, which are condensed and deprotected to obtain MJ 10813. MS (m/z): 861.3[ M +1] +.

Example 19: synthesis of MJ10814

Reference is made to the synthesis of MJ10821, except that Cbz-Ala-Ala-OH and II1 were used as starting materials, which were condensed and deprotected to give MJ 10814. MS (m/z): 875.3[ M +1] +.

Example 20: synthesis of MJ10815

Reference is made to the synthesis of MJ10821, except that Cbz-Val-Ala-OH and II1 are used as starting materials, which are condensed and deprotected to finally obtain MJ 10815. MS (m/z): 903.4[ M +1] +.

Example 21: synthesis of MJ10804

Reference is made to the synthesis of MJ10821, which differs in that Cbz-Gly-Pro-OH and II2 are selected as starting materials, and are subjected to condensation and deprotection to finally obtain MJ 10804. MS (m/z): 871.4[ M +1] +.

Example 22: synthesis of MJ10805

Reference is made to the synthesis of MJ10821, with the difference that Cbz-Ala-Pro-OH and II2 are used as starting materials, which are condensed and deprotected to finally obtain MJ 10805. MS (m/z): 885.4[ M +1] +.

Example 23: synthesis of MJ10806

Reference is made to the synthesis of MJ10821, which differs in that Cbz-Val-Pro-OH and II2 are selected as starting materials, and condensation and deprotection are carried out to finally obtain MJ 10806. MS (m/z): 913.4[ M +1] +.

Example 24: synthesis of MJ10816

Reference is made to the synthesis of MJ10821, except that Cbz-Gly-Ala-OH and II2 are used as starting materials, which are condensed and deprotected to obtain MJ 10816. MS (m/z): 845.4[ M +1] +.

Example 25: synthesis of MJ10817

Reference is made to the synthesis of MJ10821, except that Cbz-Ala-Ala-OH and II2 were used as starting materials, which were condensed and deprotected to give MJ 10817. MS (m/z): 859.4[ M +1] +.

Example 26: synthesis of MJ10818

Reference is made to the synthesis of MJ10821, except that Cbz-Val-Ala-OH and II2 are used as starting materials, which are condensed and deprotected to finally obtain MJ 10818. MS (m/z): 887.4[ M +1] +.

Example 27: synthesis of MJ10810

Reference is made to the synthesis of MJ10821, except that Cbz-Gly-Pro-OH and II4 are used as starting materials, which are condensed and deprotected to finally obtain MJ 10810. MS (m/z): 871.4[ M +1] +.

Example 28: synthesis of MJ10811

Reference is made to the synthesis of MJ10821, with the difference that starting materials Cbz-Ala-Pro-OH and II4 were selected, which were condensed and deprotected to finally obtain MJ 10811. MS (m/z): 885.4[ M +1] +.

Example 29: synthesis of MJ10812

Reference is made to the synthesis of MJ10821, which differs in that Cbz-Val-Pro-OH and II4 are selected as starting materials, and condensation and deprotection are carried out to finally obtain MJ 10812. MS (m/z): 913.4[ M +1] +.

Example 30: synthesis of MJ10822

Reference is made to the synthesis of MJ10821, except that Cbz-Gly-Ala-OH and II4 are used as starting materials, which are condensed and deprotected to obtain MJ 10822. MS (m/z): 845.4[ M +1] +.

Example 31: synthesis of MJ10823

Reference is made to the synthesis of MJ10821, except that Cbz-Ala-Ala-OH and II4 were used as starting materials, which were condensed and deprotected to give MJ 10823. MS (m/z): 859.4[ M +1] +.

Example 32: synthesis of MJ10824

Reference is made to the synthesis of MJ10821, except that Cbz-Val-Ala-OH and II4 are used as starting materials, which are condensed and deprotected to finally obtain MJ 10824. MS (m/z): 887.4[ M +1] +.

Example 33: synthesis of MJ10828

Referring to the synthesis of MJ10821, the difference is that Cbz-Trp (Boc) -Ala-OH and II3 are selected as starting materials, and the final MJ10828 is obtained by condensation, removal of tert-butoxycarbonyl protection by hydrogen chloride, and removal of benzyloxycarbonyl protection by catalytic hydrogenation. MS (m/z): 958.5[ M +1] +.

Example 34: synthesis of MJ10829

Referring to the synthesis of MJ10821, the difference is that Cbz-Tyr (tBu) -Ala-OH and II3 are selected as starting materials, and MJ10829 is finally obtained through condensation, tert-butyl protection removal by hydrogen chloride, and debenzyloxy carbonyl protection removal by catalytic hydrogenation. MS (m/z): 935.4[ M +1] +.

Example 35: synthesis of MJ10831

reference is made to the synthesis of MJ10821, which is different from the synthesis of MJ10831, in that Cbz-Asp (OBzl) -Ala-OH and II3 are selected as starting materials, and are subjected to condensation and catalytic hydrogenation to remove carbobenzoxy protection. MS (m/z): 887.4[ M +1] +.

Example 36: synthesis of MJ10832

Referring to the synthesis of MJ10821, the difference is that Cbz-Glu (OBzl) -Ala-OH and II3 are selected as starting materials, and MJ10832 is finally obtained through condensation and catalytic hydrogenation to remove carbobenzoxy protection. MS (m/z): 901.4[ M +1] +.

Example 37: synthesis of MJ10834

referring to the synthesis of MJ10821, the difference is that Cbz-Met-Ala-OH and II3 are selected as starting materials, and MJ10834 is finally obtained through condensation and Raney Ni catalytic hydrogenation debenzylation oxycarbonyl protection. MS (m/z): 903.4[ M +1] +.

Example 38: synthesis of MJ10835

Referring to the synthesis of MJ10821, the difference is that Cbz-Lys (Cbz) -Ala-OH and II3 are selected as starting materials, and MJ10835 is obtained by condensation and catalytic hydrogenation to remove carbobenzoxy protection. MS (m/z): 900.5[ M +1] +.

Example 39: synthesis of MJ10836

Referring to the synthesis of MJ10821, the difference is that Cbz-Ser (tBu) -Ala-OH and II3 are selected as starting materials, and MJ10836 is finally obtained through condensation, tert-butyl protection removal by hydrogen chloride, and debenzyloxy carbonyl protection removal by catalytic hydrogenation. MS (m/z): 859.4[ M +1] +.

Example 40: synthesis of MJ10837

Referring to the synthesis of MJ10821, the difference is that Cbz-Thr (tBu) -Ala-OH and II3 are selected as starting materials, and MJ10837 is finally obtained through condensation, tert-butyl protection removal by hydrogen chloride, and debenzyloxycarbonyl protection removal by catalytic hydrogenation. MS (m/z): 873.4[ M +1] +.

Example 41: synthesis of MJ10838

Referring to the synthesis of MJ10821, the difference is that Cbz-Cys (Trt) -Ala-OH and II3 are selected as starting materials, and MJ10838 is finally obtained through condensation, trityl protection removal by hydrogen chloride, and benzyloxycarbonyl protection removal by Raney Ni catalytic hydrogenation. MS (m/z): 875.4[ M +1] +.

Example 42: synthesis of MJ10840

referring to the synthesis of MJ10821, the difference is that Cbz-His (Trt) -Ala-OH and II3 are selected as starting materials, and the synthesis is carried out by condensation, trityl protection removal by hydrogen chloride, and debenzyloxy carbonyl protection removal by catalytic hydrogenation to finally obtain MJ 10840. MS (m/z): 909.4[ M +1] +.

Example 43: synthesis of MJ10841

Referring to the synthesis of MJ10821, the difference is that Cbz-Arg (Boc)2-Ala-OH and II3 are selected as starting materials, and MJ10841 is finally obtained by condensation, removal of tert-butoxycarbonyl protection by hydrogen chloride, and removal of benzyloxycarbonyl protection by catalytic hydrogenation. MS (m/z): 928.5[ M +1] +.

Example 44: synthesis of MJ10842

Referring to the synthesis of MJ10821, the difference is that Cbz-Lys (Cbz) -Pro-OH and II3 are selected as starting materials, and MJ10842 is finally obtained through condensation and catalytic hydrogenation to remove carbobenzoxy protection. MS (m/z): 926.5[ M +1] +.

example 45: synthesis of MJ10843

Referring to the synthesis of MJ10821, the difference is that Cbz-Ser (tBu) -Pro-OH and II3 are selected as starting materials, and MJ10843 is finally obtained through condensation, tert-butyl protection removal by hydrogen chloride, and debenzyloxy carbonyl protection by catalytic hydrogenation. MS (m/z): 885.4[ M +1] +.

Example 46: preparation of compound hydrochloride

Dissolving the compound in a mixed solvent of methanol and ethyl acetate, dropwise adding an ethyl acetate solution of hydrogen chloride, spin-drying the solvent, and washing the solid with the mixed solvent of ethyl acetate and petroleum ether for 3 times to obtain the hydrochloride of the compound MJ10801-MJ 10845.

Example 47: test for solubility of Compounds

The product is taken, ground into fine powder and operated according to the solubility of the two examples in the 2010 version of the Chinese pharmacopoeia. Accurately weighing a proper amount of a sample, adding water, placing in a water bath at 25 ℃, shaking for 30 seconds every 5 minutes, observing the dissolution condition within 30 minutes, and if no visible particles are considered to be dissolved, the comparison result of the solubility of the hydrochloride of the invention and the posaconazole is shown in table 1;

TABLE 1 results of dissolution test of the compounds

Sample (I)	Solubility (%)
		Posaconazole	<0.01％
MJ10801-MJ 10806 (hydrochloride)	>3.5％
		MJ 10807-MJ 10809 (hydrochloride)	>3.2％
MJ 10810-MJ 10818 (hydrochloride)	>3.5％
		MJ 10819-MJ 10821 (hydrochloride)	>3.2％
MJ 10822-MJ 10824 (hydrochloride)	>3.5％
		MJ 10825-MJ 10845 (hydrochloride)	>3.2％

The above results show that the compound of the present invention (hydrochloride) has more than 300 times higher water solubility than posaconazole.

Posaconazole is a white powder with low water solubility, the drug is required to have good solubility when being injected, the pH of the solution is adjusted by hydrochloric acid, the content of hydrogen chloride is not obvious when the content of hydrogen chloride is increased to 5 percent, the acid condition cannot be used for intravenous administration, and a small amount of dissolved drug can be quickly separated out under the pH buffering condition of blood.

Clinical administration used sulfobutylbetacyclodextrin sodium as a co-solvent and adjusted the pH to 2.6, but still required central intravenous administration to reduce the incidence of phlebitis. The compound can form a salt with acid due to the existence of basic groups, so that the solubility is increased, and convenience is brought to preparation.

Example 48: pharmaceutical compositions formulated with hydrochloride salts of the compounds of the invention as active ingredient

The pharmaceutical composition comprises hydrochloride of the compound, sodium sulfobutylbetacyclodextrin, disodium ethylene diamine tetraacetate and distilled water.

Example 49: comparison of rabbit ear vein vascular irritativeness

the clinically used posaconazole injection needs to be diluted and administered through a central vein, and peripheral intravenous drip cannot be used, otherwise embolic phlebitis is easily generated. The compounds of the present invention were administered in the same manner as posaconazole injection.

Preparation of posaconazole injection: 300mg of posaconazole is mixed and dissolved with 6.68g of sodium sulfobutylbetacyclodextrin and 0.003g of disodium ethylenediamine tetraacetate, and the pH value is adjusted to 2.6 by using hydrochloric acid and sodium hydroxide to prepare concentrated stock solution, wherein the final volume is 16.7 ml. The concentrated stock solution was diluted with 150ml of 0.9% sodium chloride injection to prepare posaconazole injection.

The preparation of the compound injection of the invention comprises the following steps: the compound hydrochloride of the present invention in an equimolar amount to 300mg of posaconazole was mixed with 6.68g of sulfobutylbetacyclodextrin sodium and 0.003g of disodium ethylenediaminetetraacetate to dissolve, and diluted to 16.7ml with distilled water to prepare a concentrated stock solution. The concentrated stock solution is diluted with 150ml of 0.9% sodium chloride injection to prepare the compound injection.

Every 3 rabbits were divided into left and right ears, 2 injections were administered via the rabbit ear vein, and were administered by intravenous drip at a low rate of 7.8ml/20min for 3 days, and animals were sacrificed on day 4.

The rabbit ears are obtained by taking materials conventionally, and the proximal, middle and distal sections are taken from the ear tips to the ear roots. From the tip to the root of the ear, a proximal section is 1.3cm from the needle insertion site, a middle section is about 2.6cm from the needle insertion site, and a distal section is 4.0cm from the needle insertion site. The material selection range can be properly enlarged according to the needle insertion condition. Finally, dehydrating, embedding, staining and performing optical microscopy, wherein all morphological changes are marked as +/-plus or minus, plus or minus respectively, and represent slight, mild, moderate and severe, and are respectively scored as 1, 2, 3 and 4, and non-pathological change is marked as < - > and is scored as 0; the results of the rabbit ear intravenous drip test using the invention and posaconazole injection are shown in table 2;

TABLE 2 results of intravenous drip test of posaconazole injection solution on rabbit ear

Note: all morphological changes are marked "±" + + + ", respectively for mild, moderate, severe, no lesion as" - ", and deletions as" none ".

The compound is administrated in the same way as the posaconazole injection, and the ear veins of the posaconazole group show black hair and involve vein branches by visual observation; the rabbit ear vein of the compound of the invention showed red color and no significant change in the venous branches.

Example 50: biotransformation assay of Compounds in human plasma

The compounds of the invention need to be converted in vivo to compounds of formula II for pharmaceutical action, so the degree of in vivo biotransformation is directly related to the potency. Dissolving a test compound by using methanol to prepare a solution with the concentration of 10mg/ml, taking 3ul, adding the solution into 300ul of plasma, incubating for 2 hours at 37 ℃, taking 100ul of incubated plasma, adding 900ul of methanol, vortexing for 3min, centrifuging for 3min at 10000 rpm, feeding 20ul of supernate, analyzing by using high performance liquid chromatography, and calculating the conversion rate by using an area normalization method, namely the peak area of the compound shown in the formula II/(the peak area of the compound shown in the formula II + the peak area of the compound shown in the invention). The results are shown in Table 3:

TABLE 3 conversion of the compounds of the invention at human plasma incubation for 2h

Compound (I)	MJ10801-10812	MJ108013-10841	MJ108042-10843	MJ108044-10845
					Conversion rate	95％	～63％	99％	～60％

Example 51: antifungal infection tests Using the Compounds of the invention

48 male Kunming mice (20-24 g) were divided into 6 groups of 8 mice each, namely a normal saline group, a posaconazole group and compound MJ10821, MJ10835, MJ10839 and MJ10842 groups. After 4 hours, each mouse was injected intravenously (0.1 ml/mouse) with an infection solution containing Candida albicans (isolated clinically), and after intraperitoneal injection (0.2ml/10g body weight) of physiological saline and each test solution (the vehicle was physiological saline containing sodium sulfobutylbetacyclodextrin and disodium ethylenediaminetetraacetate, which was prepared according to the method of example 49 and further diluted with physiological saline), once a day, for three consecutive days, and after four days, the number of surviving animals was counted, and the results are shown in Table 4.

TABLE 4 results of antifungal infection test

as can be seen from the data in Table 4, the survival rate of mice is higher than that of posaconazole group when the drug is used for antifungal infection tests, and the antibacterial effect of the drug is obviously better than that of posaconazole, so that the product has better effect on antifungal infection.

In conclusion, the medicine provided by the invention can prevent the stimulation of too low pH of the injected medicine to blood vessels during intravenous injection, and can ensure that the antibacterial effect of the medicine is in a better level.

As noted above, while the present invention has been shown and described with reference to certain preferred embodiments, it is not to be construed as limited thereto. Various changes in form and detail may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims (4)

1. A compound of formula I, or a pharmaceutically acceptable acid salt thereof:

Wherein:

x1 is fluoro or chloro;

X2 is fluoro or chloro;

R1 is a dipeptide acyl or polypeptide acyl group wherein at least one amino acid acyl group is formed from a natural amino acid; the compound is selected from the group of the following structures:

or a pharmaceutically acceptable acid salt thereof.

2. A pharmaceutical composition comprising as an active ingredient a therapeutically effective amount of a compound of formula I according to claim 1 or a pharmaceutically acceptable acid salt thereof.

3. Use of a compound of formula I according to claim 1 or a pharmaceutically acceptable acid salt thereof or a pharmaceutical composition according to claim 2 for the manufacture of a medicament for the prophylaxis or treatment of fungal infections.

4. Use according to claim 3, characterized in that: the fungus is selected from Candida, Histoplasma capsulatum, Sedum cerevisia, Zygomycetes bipolaris, Fusarium, yeast, and NonC albicans, Cryptococcus neoformans or Aspergillus with fluconazole resistance.