Amphotericin B peptide derivative and preparation method thereof
A technology of amphotericin and compounds, applied in the field of medicine, can solve the problems of increasing treatment dosage, high price and high cost
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Embodiment 1
[0085] Example 1: Preparation and purification of DMR005
[0086] Hydrophilic peptide structure: Fmoc-AEEAc-AEEAc-AEEAc-AEEAc–AEEAc-OH
[0087] DMR005 structure:
[0088]
[0089] (1) Materials and reagents
[0090] 2-CTC resin, substitution value 0.945mmol / g.
[0091] Amino acid: Fmoc-AEEAc-OH
[0092] Synthetic reagents: HATU, DMF, DCM, DIEA, piperidine.
[0093] (2) Instruments
[0094] CS-BIO peptide synthesizer, Waters600 semi-preparative high performance liquid chromatograph, Beckman centrifuge, Buchi rotary steamer.
[0095] (3) Operation steps (take 1g resin as an example)
[0096] a. Solid-phase chemical synthesis of peptides
[0097] Weigh 1.00g of 2-CTC resin, put it in the peptide synthesizer reactor, add 10mLDCM, soak for 1h, weigh 2-3 times the amount of Fmoc-AEEAc-OH and draw 4-6 times the amount of DIEA, add 10mLDCM to dissolve it , Put it into the reactor and proceed with the reaction. The reaction temperature is room temperature (25°C and above, otherwise the reaction time ...
Embodiment 2
[0122] Example 2: Contain n NH 2 -AEEAc-OH Preparation and Purification of Amphotericin B Peptide Derivatives
[0123] Amino acid sequence: Fmoc-(AEEAc) n -OH; n=1-20, n is an integer, the product structure is as follows:
[0124]
[0125] Synthetic NH 2 -(AEEAc) n -Amphotericin B peptide derivative of AMB, the synthesis method refers to compound DMR005, namely ① Fmoc-(AEEAc) is synthesized by solid phase synthesis n -OH; ②Preparation of Fmoc-(AEEAc) n -Osu; ③Fmoc-(AEEAc) n -Osu modifies the amino group of amphotericin B, piperidine removes Fmoc, and finally obtains NH 2 -(AEEAc) n -Crude amphotericin B peptide derivative of AMB. The crude product was purified by RP-HPLC; according to the general structure of the compound [Ⅱ], Table 3 lists the abbreviations and relative molecular weights of the compounds:
[0126] Table 3 Compound abbreviations and relative molecular masses
[0127] namenR Relative molecular mass NH 2 -AEEAc-AMB
Embodiment 3
[0128] Example 3: Determination of the solubility of amphotericin B peptide derivatives
[0129] For the determination of the solubility of amphotericin B and amphotericin B peptide derivatives, in the patent of the present invention, the solubility in water was measured. The results are shown in Table 4:
[0130] Table 4 Solubility measurement results
[0131] Compound Solubility (mg / mL) AMB<0.001
[0132] The results show that AMB is almost insoluble in water; NH 2 -AEEAc-AMB has lower solubility, DMR005 obviously improves water solubility; with NH 2 -(AEEAc) n The increase of n in -OH significantly increases the solubility of the amphotericin B peptide derivative. But at the same time, the antibacterial activity was significantly reduced (see Table 5 for the results of antibacterial activity), and even no antibacterial activity. When n is 3-7, solubility and antibacterial activity are in a good balance.
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