CN104557743A - Candida albicans-resistant medicines with unsaturated fatty acid structure and preparation method of candida albicans-resistant medicines - Google Patents

Abstract

The invention discloses a method for rapidly preparing and screening an anti-candida albicans drug containing an unsaturated fatty acid structure. The structure of this class anti-candida drug analogue or derivative is represented by general formula (II) Said that the drug properties were screened through antibacterial experiments. The invention discloses a preparation method and a screening method of a synthetic drug, which can not only reduce the development cost of new drugs, but also effectively improve the efficiency of new drug detection. Wherein, R ₁ represents one of linear alkyl, terminal hydroxyl or mercapto-substituted alkyl, oligoethylene glycol or 1,4-alkylpiperazine, R ₂ is hydroxyl, amino, methoxy A sort of.

Description

A kind of anti-candida albicans drug containing unsaturated fatty acid structure and preparation method thereof

technical field

The invention belongs to the field of drug design and drug screening, and specifically relates to the field of preparation of a class of anti-candida albicans drugs.

Background technique

Candida albicans (Candida albicans) is a common opportunistic fungus in humans and one of the main pathogens of clinically acquired infections. Under normal circumstances, it exists in the form of symbiotic bacteria on the skin, oral cavity, gastrointestinal tract and mucosal surface of the human body. When the human immune system is normal, Candida albicans, as a symbiotic bacterium, maintains a balance with other microbial flora in the body and will not cause disease infection. However, when immunity is weakened or antibacterial drugs are overused, it may rapidly expand on the mucosal surface and cause Candidiasis. The most common clinically are oral candidiasis and candidal vaginitis. In addition, Candida can enter endothelial cells through tissue damage, or form biofilms and colonize built-in medical devices, such as various catheters, artificial joints, dentures, etc., and thus enter the blood, leading to severe candidemia.

With the extensive use of clinical antibiotics, anticancer drugs and immunosuppressants, the incidence of candidiasis is showing a sharp upward trend, and the threat to the human body is increasing, especially for severe patients and bone marrow transplant recipients. Candida infections can be life-threatening. Therefore, the effective treatment of candidiasis is particularly urgent. In the past ten years, the research and development of antifungal drugs has been continuously developed, and various drugs have been used in clinic one after another, and good results have been achieved. For example, the application of antifungal drugs such as amphotericin B has saved the lives of countless patients with fungal diseases. life. But at the same time, serious problems have arisen. Long-term use can lead to the fatal problem of enhanced drug resistance of bacterial strains. Therefore, it is particularly urgent and important to find new antifungal drugs that are natural, low-toxic, highly effective, and especially capable of overcoming clinical drug resistance, and to propose new control strategies.

Triazole antifungal drugs are a class of synthetic antifungal drugs developed in recent years. Their mechanism of action is to inhibit the cytochrome P450 that catalyzes lanosterol 14α-demethylation to generate ergosterol, resulting in the lack of ergosterol, thus making the cell membrane Structural and functional disappearance and destruction. In recent years, research on this type of drug has developed rapidly, from early miconazole drugs such as ketoconazole, econazole, etc. to triazole drugs, such as itraconazole, fluconazole, etc., but some Triazoles have long-term tolerance and considerable side effects.

In recent years, some natural small molecular compounds that have the activity of regulating or inhibiting the morphological transformation of Candida albicans have attracted much attention because they are expected to become potential new antifungal drugs. Oh et al. reported in 2001 a form-regulating substance (ARS) secreted by Candida albicans --- farnesic acid, which does not affect the growth and reproduction of Candida albicans, but can regulate the morphological transformation of Candida albicans and inhibit hyphae grow. A chemical signaling molecule of Candida albicans quorum effect --- farnesol can also effectively inhibit the growth of Candida albicans hyphae. In addition, 3-oxo-lauroyl homoserine cyclic lactone, a quorum-sensing signaling molecule produced from Pseudomonas aeruginosa, can interfere with the morphological transition of Candida albicans at high concentrations. In recent years, two new signaling molecules --- α, β-cis-unsaturated fatty acid cis-11-methyldodeca-2-enoic acid ( DSF) and cis-dodeca-2-enoic acid (BDSF), can regulate and inhibit the activity of Candida albicans morphological transformation and reduce its pathogenicity. Although the mechanism of action of these small molecules is still poorly understood, their ability to regulate the morphological transformation of Candida albicans has brought new enlightenment to our treatment of fungal infections. Therefore, it is particularly important to design and synthesize a series of analogs according to the structural characteristics of these small molecular compounds and triazoles, and to study their inhibitory activity against Candida albicans.

However, the synthesis of DSF drugs requires at least three steps (such as III), the selected raw material fatty acid needs to be obtained by industrial production, methyllithium is expensive, and the bromination reagent liquid bromine is more toxic and seriously pollutes the environment. Not only the cost is high, but also The operation is cumbersome. The synthesis method selected by the present invention has short route, simple operation, and cheap and easy-to-obtain raw materials such as copper sulfate, which greatly reduces the development cost of new drugs.

Contents of the invention

Technical problem: The purpose of the present invention is to design and synthesize a new class of drugs, provide its preparation process, and form a rapid drug screening method.

Technical scheme: a class of anti-Candida albicans medicine comprising unsaturated fatty acid structure of the present invention, its structure can be represented by formula (II):

Wherein, R ₁ represents any one of linear alkyl, terminal hydroxyl or mercapto-substituted alkyl, oligoethylene glycol or 1,4-alkylpiperazine, R ₂ is hydroxyl, amino, methoxy of any kind.

The unsaturated fatty acid (ester, amide) in the structure of formula (II) can play a bacteriostatic effect in organisms; the imidazole ring is an effective bacteriostatic pharmacophore.

The preparation method comprises the following synthetic steps:

Step i. Under the protection of nitrogen, the halide R ₁ X, the catalyst and sodium azide are reacted in an organic solvent at 20-100°C for 12-24 hours in a sealed manner to obtain the azide compound R ₁ N ₃ shown in formula (I);

Step ii. Sodium ascorbate is dissolved in the solvent, and the aqueous solution of hydrated divalent copper ion salt, propioloyl R ₂ compound and azide compound R ₁ N ₃ are sequentially added, and the reaction is sealed at room temperature under nitrogen protection for 5 to 10 hours, and separation and purification are carried out. Obtain the target compound shown in formula (II).

R described in step _i represents one of straight-chain alkyl, terminal hydroxyl or mercapto-substituted alkyl, oligoethylene glycol or 1,4-alkylpiperazine; said X represents is Cl, Br, I or trifluoromethanesulfonyl, the catalyst is potassium iodide, and the organic solvent is tetrahydrofuran, N,N-dimethylformamide, one of dimethyl sulfoxide; step ii Described hydrated divalent copper ion salt is copper sulfate pentahydrate or cupric chloride dihydrate; Described R ₂ is a kind of in hydroxyl, amino, methoxy group; Described solvent is methanol, ethanol or tert-butanol Mixture with water volume ratio 1:1 or 2:1.

Beneficial effect

1. The drug structure proposed by the present invention not only includes a bactericidal structure, but also an antibacterial structure, and has a certain effect on the treatment of Candida albicans.

2. The medicine proposed by the present invention has a short synthesis route and cheap and easy-to-obtain raw materials, which can greatly reduce the development cost of new medicines.

3. The method of the present invention has a short route and simple operation. A series of analogs or derivatives of antifungal drugs with different structures are synthesized, and drug properties are screened through antibacterial experiments to form a method for rapidly screening antifungal drugs.

Description of drawings

Fig. 1. Candida albicans (C.albicans) hyphal growth status (control, mycelial state, shows that Candida albicans is in active state);

Figure 2.30 μ M antifungal drugs under the action of Candida albicans (C.albicans) hyphae growth (yeast state, indicating that Candida albicans is in a silent state);

Figure 3. The growth of Candida albicans (C.albicans) mycelia under the action of 300 μM antifungal drugs (yeast state, indicating that Candida albicans is in a state of population silence).

Figure 4. The growth curve of Candida albicans (C.albicans) under the action of 100 μM antifungal drugs for 0-24h.

Detailed ways

In order to better understand the content of the present invention, the technical solutions of the present invention will be further described below through specific examples. However, these implementation examples do not limit the present invention.

Example 1:

Step 1. In a two-necked round bottom flask (100mL), add sodium azide (0.975g, 15mmol), potassium iodide (0.083g, 0.5mmol), nitrogen protection, add dimethyl sulfoxide 15mL, nitrogen protection, at 50°C Stir. Then, bromooctane (1.92 g, 10 mmol) was added to the reaction system. After the dropwise addition, continue to stir at 50°C for 10h. The reaction was quenched with ice water, extracted with petroleum ether, the organic phase was washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain 1.3 g of octane azide with a yield of 87%.

Step 2. Add sodium ascorbate (0.17g, 0.85mmol) into a two-neck round bottom flask (100mL) under nitrogen protection. Then 3 mL of ethanol was added. Nitrogen was blown, and 3 mL of an aqueous solution of copper sulfate (0.1 g, 4 mmol) was added, and then propiolic acid (0.73 g, 10 mmol) and octane azide (1.35 g, 8.7 mmol) were added dropwise to react for 5 h. The reaction was quenched by adding ice water. Acidify with dilute hydrochloric acid, extract with ethyl acetate, concentrate, and separate and purify by silica gel column chromatography. 1.27 g of product (1) was obtained, yield 65%. ¹ H NMR (400MHz, DMSO) δ (ppm): 11.0 (s, 1H), 8.67 (s, 1H), 4.37 (t, J = 7.1Hz, 2H), 1.90–1.73 (m, 2H), 1.34–1.10 (m,10H),0.83(t,J=6.9Hz,3H).

Example 2-Example 6, method is the same as Example 1, and the specific experimental consumption is as follows:

Step 1

R ₁ R ₁ Br (g, mmol) NaN ₃ (g, mmol) KI (g, mmol) DMSO (mL) R ₁ N ₃ (g) Yield(%) 2 C ₆ H ₁₃ 1.64,10 0.975,15 0.083,0.5 15 0.9 70 3 C ₆ H ₁₂ OH 1.80,10 0.975,15 0.083,0.5 15 0.9 63 4 C ₁₂ H ₂₅ 2.50,10 0.975,15 0.083,0.5 15 1.4 70 5 C ₈ H ₁₇ 1.92,10 0.975,15 0.083,0.5 15 1.4 90 6 C ₈ H ₁₇ 1.92,10 0.975,15 0.083,0.5 15 1.3 84

Step 2

characterizing data

Product 2:

¹ H NMR (400MHz, DMSO) δ (ppm): 11.0 (s, 1H), 8.68 (s, 1H), 4.45–4.31 (m, 2H), 1.87–1.73 (m, 2H), 1.22 (pd, J =8.0,5.0Hz,6H),0.82(t,J=7.0Hz,3H).

Product 3:

¹ H NMR (400MHz, DMSO) δ (ppm): 11.2 (s, 1H), 8.27 (s, 1H), 4.34 (t, J = 14.1, 7.2Hz, 2H), 1.80 (m, J = 31.2, 23.7 Hz,2H),1.43(m,J=45.5,6.9Hz,1H),1.21(d,J=8.8Hz,13H),0.82(m,J=8.0,5.6Hz,3H).

Product 4:

¹ H NMR (400MHz, DMSO) δ (ppm): 11.3 (s, 1H), 8.27 (s, 1H), 4.34 (t, J = 14.1, 7.1 Hz, 2H), 1.76 (m, J = 14.4, 7.1 Hz, 2H), 1.21(d, J=6.3Hz, 18H), 0.82(t, J=6.7Hz, 3H).

Product 5:

¹ H NMR (400MHz, DMSO) δ (ppm): 8.67 (s, 1H), 4.37 (t, J = 7.1Hz, 2H), 4.26 (m, 2H), 1.90–1.73 (m, 2H), 1.34– 1.10(m,10H),1.09(t,3H)0.83(t,J＝6.9Hz,3H).

Product 6:

¹ H NMR (400MHz, DMSO) δ (ppm): 8.79 (s, 2H), 8.67 (s, 1H), 4.37 (t, J = 7.1Hz, 2H), 1.90–1.73 (m, 2H), 1.34–1.10 (m,10H),0.83(t,J=6.9Hz,3H).

Claims (4)

1. a class of anti-candida albicans medicine comprising unsaturated fatty acid structure, is characterized in that this class medicine is the compound of following formula (II): Wherein, R ₁ represents any one of linear alkyl, terminal hydroxyl or mercapto-substituted alkyl, oligoethylene glycol or 1,4-alkylpiperazine, and R ₂ is hydroxyl, amino, methoxy any of the bases. 2. the preparation method of the anti-candida albicans medicine comprising unsaturated fatty acid structure described in claim 1, is characterized in that, the preparation method of this class medicine comprises the following synthetic steps: Step i. Under the protection of nitrogen, the halide R ₁ X, the catalyst and sodium azide are reacted in an organic solvent at 20-100° C. for 12-24 hours in a sealed manner to obtain the azide R ₁ N ₃ compound shown in formula (I); Step ii. Sodium ascorbate is dissolved in the solvent, and the aqueous solution of hydrated divalent copper ion salt, propioloyl R ₂ compound and azide compound R ₁ N ₃ are sequentially added, and the reaction is sealed at room temperature under nitrogen protection for 5 to 10 hours, and separation and purification are carried out. Obtain the target compound shown in formula (II). 3. the preparation method of a kind of anti-candida albicans medicine that comprises unsaturated fatty acid structure according to claim 2, it is characterized in that the R described in step _i represents that straight-chain alkyl, terminal hydroxyl or sulfhydryl replace Any one of alkyl, oligoethylene glycol or 1,4-alkylpiperazine; said X represents Cl, Br, I or trifluoromethanesulfonyl, and said catalyst is potassium iodide, The organic solvent is one of tetrahydrofuran, N,N-dimethylformamide and dimethyl sulfoxide. 4. the preparation method of a kind of anti-Candida albicans medicine that comprises unsaturated fatty acid structure according to claim 2, it is characterized in that the hydration divalent copper ion salt described in step ii is copper sulfate pentahydrate or dihydrate chlorination Copper; the _R2 is one of hydroxyl, amino, and methoxy; the solvent is a mixture of methanol or ethanol or tert-butanol, and the volume ratio of solvent to water is 1:1 or 2:1.