CN101747410B - Aminoacyl tyrosyl tryptophan tripeptide with easing pain activity and preparation method and application thereof - Google Patents

Abstract

The invention discloses an aminoacyltyrosyl tryptophan tripeptide with analgesic activity, a preparation method and an application. The structural formula of aminoacyl tyrosyl tryptophan tripeptide of the present invention is AA-Tyr-Trp, wherein, AA is selected from alanine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, Isoleucine, Lysine, Leucine, Methionine, Asparagine, Proline, Glutamine, Arginine, Serine, Threonine, Valine, Tryptophan, or Tyramine acid residues. Animal experiments show that the compound of the present invention has excellent analgesic activity and is a clinical potential analgesic.

Description

Aminoacyltyrosyltryptophan tripeptide with analgesic activity, preparation method and application

technical field

The present invention relates to tripeptides, in particular to aminoacyltyrosyl tryptophan tripeptides with analgesic activity and a preparation method, and also to the use of the aminoacyltyrosyl tryptophan tripeptides in preparing analgesic drugs , belonging to the field of biomedicine.

Background technique

Pain is not only a complication of many diseases, but has been recognized as a class of diseases in itself. At the disease level, the loss of quality of life caused by pain of all kinds affects the greatest number of people. Since effective analgesics are often associated with dependence, the development of new dependence-free analgesics is of great value. In the study of peptides, the inventors accidentally discovered that tryptophan-containing oligopeptides usually have analgesic activity. According to this clue, the present invention discloses a class of aminoacyltyrosyltryptophan tripeptide analgesic.

Contents of the invention

One of the objectives of the present invention is to provide a class of aminoacyltyrosyltryptophan tripeptides with analgesic activity.

The second object of the present invention is to provide a method for preparing the above-mentioned aminoacyltyrosyltryptophan tripeptide.

The third object of the present invention is to provide a pharmaceutical composition capable of analgesia.

Above-mentioned purpose of the present invention is achieved through the following technical solutions:

A class of aminoacyltyrosyltryptophan tripeptides with analgesic activity, the structure of which is shown in general formula I:

AA-Tyr-Trp I

Wherein, AA is selected from alanine residues, aspartic acid residues, glutamic acid residues, phenylalanine residues, glycine residues, histidine residues, isoleucine residues, lysine residues, Acid residues, leucine residues, methionine residues, asparagine residues, proline residues, glutamine residues, arginine residues, serine residues, threonine residues , valine residues, tryptophan residues or tyrosine residues.

A method for preparing the above-mentioned aminoacyltyrosyltryptophan tripeptide with analgesic activity, comprising:

(1) coupling tert-butoxycarbonyl-protected tyrosine and tryptophan benzyl ester to synthesize tert-butoxycarbonyl tyrosyl tryptophan benzyl ester;

(2) removing the tert-butoxycarbonyl group of tert-butoxycarbonyl tyrosyl tryptophan benzyl ester to obtain tyrosyl tryptophan benzyl ester;

(3) Coupling tyrosyl tryptophan benzyl ester with tert-butoxycarbonyl-protected amino acid to obtain tert-butoxycarbonyl aminoacyl tyrosyl tryptophan benzyl ester;

(4) hydrogenolysis and debenzylation of tert-butoxycarbonylaminoacyl tyrosyl tryptophan benzyl ester to obtain tert-butoxycarbonyl aminoacyl tyrosyl tryptophan;

(5) Remove tert-butoxycarbonyl from tert-butoxycarbonyl aminoacyl-tyrosyl-tryptophan to obtain the aminoacyl-tyrosyl-tryptophan tripeptide of general formula I.

Among them, in the step (1), preferably in the presence of dicyclohexylcarbodiimide, N-hydroxybenzotriazole, N-methylmorpholine and anhydrous tetrahydrofuran, the tyrosine and tryptophan with tert-butoxycarbonyl protection Acid benzyl ester coupling, synthesis of tert-butoxycarbonyl tyrosyl tryptophan benzyl ester;

In step (2), it is preferable to add tert-butoxycarbonyl tyrosyl tryptophan benzyl ester to the hydrogen chloride/ethyl acetate solution to remove the tert-butoxycarbonyl group to prepare tyrosyl tryptophan benzyl ester;

In step (3), preferably in the presence of dicyclohexylcarbodiimide, N-hydroxybenzotriazole, N-methylmorpholine and anhydrous tetrahydrofuran, tyrosyl tryptophan benzyl ester and tert-butoxycarbonyl protection The amino acid of tert-butoxycarbonyl is coupled to obtain tert-butoxycarbonyl aminoacyl tyrosyl tryptophan benzyl ester; the amino acid protected by tert-butoxycarbonyl is preferably: alanine protected by tert-butoxycarbonyl, tert-butoxycarbonyl-protected monobenzyl aspartate, tert-butoxycarbonyl-protected monobenzyl glutamate, tert-butoxycarbonyl-protected phenylalanine, tert-butoxycarbonyl-protected glycine, tert-butoxycarbonyl-protected histidine , tert-butoxycarbonyl-protected isoleucine, tert-butoxycarbonyl-protected di-tert-butoxycarbonyl-protected lysine, tert-butoxycarbonyl-protected leucine, tert-butoxycarbonyl-protected methionine, tert-butoxy Carbonyl-protected asparagine, tert-butoxycarbonyl-protected glutamine, tert-butoxycarbonyl-protected nitroarginine, tert-butoxycarbonyl-protected serine, tert-butoxycarbonyl-protected threonine, tert-butoxy Carbonyl-protected valine, tert-butoxycarbonyl-protected tryptophan or tert-butoxycarbonyl-protected tyrosine;

In step (4), preferably in the presence of palladium carbon, tert-butoxycarbonylaminoacyl tyrosyl tryptophan benzyl ester is debenzylated by hydrogenolysis to prepare tert-butoxycarbonyl aminoacyl tyrosyl tryptophan;

In step (5), it is preferable to remove tert-butoxycarbonyl from tert-butoxycarbonyl aminoacyl tyrosyl tryptophan in hydrogen chloride/ethyl acetate solution to prepare the aminoacyl tyrosyl tryptophan tripeptide of general formula I.

The analgesic activity of the compound of the present invention is evaluated on a mouse tail flick model, and the experimental results show that the compound of the general formula I of the present invention has excellent analgesic activity and can be used as an analgesic clinically. On this basis, another object of the present invention is to provide a pharmaceutical composition with analgesic activity, which consists of a therapeutically effective dose of the compound of general formula I of the present invention and a pharmaceutically acceptable excipient agent or adjuvant, that is, after compounding an effective amount of the compound of the present invention with a pharmaceutically acceptable carrier or diluent, it is prepared into any suitable pharmaceutical composition according to the conventional preparation methods in the art. Usually the composition is suitable for oral administration and injection administration, and other administration methods are also suitable. The composition can be in the form of tablets, capsules, powders, granules, lozenges, suppositories, or liquid preparations such as oral liquids. According to different administration methods, the pharmaceutical composition of the present invention may contain 0.1%-99% by weight, preferably 10-60% by weight of the compound of the present invention.

Description of drawings

Figure 1 is a synthetic route diagram of the compounds of the present invention; (i) DCC, HOBt, NMM, THF; (ii) 4NHCl/EtOAc, 0°C, 30min; (iii) H ₂ /Pd; Boc-AA selected from tert-butoxy Carbonyl-protected alanine, monobenzyl aspartate, monobenzyl glutamate, phenylalanine, glycine, histidine, isoleucine, di-tert-butoxycarbonyl-protected lysine, leucine amino acid, methionine, asparagine, glutamine, nitroarginine, serine, threonine, valine, tryptophan, or tyrosine.

Detailed ways

The present invention will be further described below in conjunction with specific embodiments, and the advantages and characteristics of the present invention will become clearer along with the description. However, these embodiments are only exemplary and do not constitute any limitation to the scope of the present invention. Those skilled in the art should understand that the details and forms of the technical solutions of the present invention can be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the protection scope of the present invention.

Embodiment 1 prepares Ala-Tyr-Trp (AYW)

1) Preparation of Boc-Tyr-Trp-OBz1

Dissolve Boc-Tyr-OH (20.00g, 71.17mmol) and HOBt (9.60g, 71.17mmol) in anhydrous THF at room temperature, adjust the pH value to 8-9 with NMM under ice cooling, add DCC (14.66g, 71.17mmol), a large amount of dicyclohexyl urea (DCU) was precipitated by stirring at 0°C. After 15-20 minutes, H ₃ PO ₄ ·Trp-OBz1 (23.25g, 59.31mmol) was added to the reaction solution. After half an hour, the ice bath was removed and stirred at room temperature. After 12 hours, the stirring was stopped, DCU was filtered, and THF was spin-dried as much as possible to obtain a yellow viscous liquid. Dissolved in 300ml ethyl acetate, washed with saturated sodium bicarbonate (200ml×3), saturated saline (200ml×2), 5% KHSO ₄ (200ml×3), saturated saline (200ml×2), anhydrous Na2SO4 dried. Concentrate under reduced pressure to remove the solvent to obtain a yellow viscous liquid, add diethyl ether to dry under reduced pressure, and repeat several times to obtain 36.473 g (92%) of a light yellow solid, which is directly used in the next step without separation by column chromatography. ESI ⁺ -MS(m/e)558[M+H] ⁺ .

2) Preparation of HCl Tyr-Trp-OBz1

Boc-Tyr-Trp-OBz1 (10.00 g, 17.95 mmol) was dissolved in anhydrous ethyl acetate, and 100 ml HCl/EtOAc (4N) was added at 0°C. After 0.5 hours the reaction was complete. Petroleum ether was added, and a large amount of powdery white solid was precipitated. After filtering, the filter residue was washed with ether (50ml×3). The filtrate was concentrated to dryness under reduced pressure to obtain 8.42 g (95%) solid, which was put into the next reaction. ESI ⁺ -MS(m/e)458[M+H]+.

3) Preparation of Boc-Ala-Tyr-Trp-OBz1

Dissolve Boc-Ala (403mg, 2.13mmol), HOBt (280mg, 2.07mmol) in anhydrous dimethylformamide (DMF), adjust the pH value to 8-9 with NMM at 0°C, add anhydrous DCC (483 mg, 2.13 mmol) dissolved in DMF was reacted for 20 minutes. Compound HCl.Tyr-Trp-OBz1 (1.0 g, 2.07 mmol) was then added. After 0.5 hour, the ice bath was removed and stirred at room temperature. After 12 hours, the stirring was stopped, DCU was filtered, and the filtrate was concentrated under reduced pressure to obtain a yellow viscous liquid. The residue was dissolved in 50ml of ethyl acetate and washed successively with saturated sodium bicarbonate (30ml×3), saturated brine (30ml×2), 5% KHSO ₄ aqueous solution (30ml×3), and saturated brine (30ml×2) , dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 973 mg (80%) of the title compound as a yellow solid. ESI ⁺ -MS(m/e)629[M+H] ⁺ .

4) Preparation of Ala-Tyr-Trp

Boc-Ala-Tyr-Trp-OBz1 (500 mg) was dissolved in methanol, 15% Pd/C was added, hydrogen was passed through for 48 hours, palladium carbon was filtered off, and the filtrate was concentrated under reduced pressure. Add 5ml of 4N HCl/EtOAc to the residue, stir at 0°C for 0.5 hours, filter, and concentrate the filtrate under reduced pressure. The residue was washed with anhydrous ether (30ml×3). Obtained 240 mg (79%) of the title compound as a colorless powder. Mp.122-123C; [α] _D ²⁵ =16.6 (c=1.23, CH ₃ OH), ESI ⁺ -MS (m/e) 439 [M+H] ⁺ ; ¹ HNMR (BHSC-300Hz, DMSO-d ₆ ), 10.82(s, 1H), 8.37(d, J=8.0Hz, 1H), 8.08(d, J=7.5Hz, 1H), 7.58(d, J=7.5Hz, 1H), 7.31(d, J=7.5Hz, 1H), 6.98(m, 5H), 6.64(d, J=8.4, 2H), 3.55(q, J=6.9Hz, J=14.1Hz, 1H), 3.10(m, 4H), 2.09 (s, 1H), 1.20 (d, J=7.5Hz, 3H).

Example 2 Preparation of Cys-Tyr-Trp (CYW)

1) Preparation of Boc-Asp(OBz1)-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 870 mg (79%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 763 [M+H] ⁺ .

2) Preparation of Asp-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 100 mg (60%) of the title compound as a pale yellow powder. Mp: 189-191C; [α] _D ²⁵ =38.7 (c=1.08, CH ₃ OH); ESI ⁺ -MS (m/e) 483 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 10.83(s, 1H), 8.35(d, J=8.0Hz, 1H), 7.98(d, J=7.5Hz, 1H), 7.56(d, J=7.5Hz, 1H), 7.32(d, J=7.5Hz, 1H), 7.19(s, 1H), 7.03(m, 4H), 6.62(d, J=8.4Hz, 2H), 4.34(m, 2H), 3.72(m, 1H), 3.22( m, 4H), 2.77 (d, J=6.5Hz, 2H), 2.09 (s, 1H).

Example 3 Preparation of Glu-Tyr-Trp (EYW)

1) Preparation of Boc-Glu(OBz1)-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 760 mg (79%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 777 [M+H] ⁺ .

2) Preparation of Glu-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 242 mg (88%) of the title compound as a pale yellow powder. Mp: 154-155C; [α] _D ²⁵ =52.6 (c=1.21, CH ₃ OH); ESI ⁺ -MS (m/e) 497 [M+H] ⁺ ; ¹ HNMR (BHSC-300Hz, DMSO-d ₆ ), 10.87(s, 1H), 9.24(s, 2H), 8.57(d, J=8Hz, 1H), 8.47(d, J=7.5Hz, 1H), 7.55(d, J=7.5Hz, 1H ), 7.35(d, J=7.5Hz, 1H), 7.18(s, 1H), 7.08(m, 4H), 6.65(d, J=8.4Hz, 2H), 4.50(m, 2H), 3.74(m , 1H), 2.98(m, 2H), 2.75(m, 2H), 2.40(d, J=7Hz, 2H), 2.10(s, 1H), 1.98(m, 2H).

Example 4 Preparation of Phe-Tyr-Trp (FYW)

1) Preparation of Boc-Phe-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 580 mg (47%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 727 [M+Na] ⁺ .

2) Preparation of Phe-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 308 mg (69%) of the title compound as a colorless powder. Mp: 148-149C; [α] _D ²⁵ =1.3 (c=1.08, CH ₃ OH); ESI ⁺ -MS (m/e) 514.9 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 12.68(s, 1H), 10.93(s, 1H), 9.25(s, 1H), 8.83(d, J=8Hz, 1H), 8.81(d, J=7.5Hz, 1H), 8.11(s , 2H), 7.57(d, J=7.5Hz, 1H), 7.36(d, J=7.5Hz, 1H), 7.21(m, 8H), 7.08(d, J=8.4Hz, 2H), 6.65(d , J=8.4Hz, 2H), 4.55(m, 2H), 4.00(s, 1H), 3.35(d, J=7Hz, 2H), 3.15(m, 4H).

Example 5 Preparation of Gly-Tyr-Trp (GYW)

1) Preparation of Boc-Gly-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 990 mg (88%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 634 [M+Na] ⁺ .

2) Preparation of Gly-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 320 mg (89%) of the title compound as a pale yellow powder. Mp: 164-167C; [α] _D ²⁵ =19.6 (c=1, CH ₃ OH); ESI ⁺ -MS (m/e) 425.3 [M+H] ⁺ ; ¹ HNMR (BHSC-300Hz, DMSO-d ₆ ), 12.68(s, 1H), 10.94(s, 1H), 9.24(s, 2H), 8.57(d, J=8.7Hz, 1H), 8.51(d, J=7.5Hz, 1H), 8.05( s, 2H), 7.56(d, J=7.5Hz, 1H), 7.35(d, J=7.5Hz, 1H), 7.19(s, 1H), 7.02(m, 2H), 6.95(d, J=8.4 Hz, 2H), 6.63(d, J=8.4Hz, 2H), 4.53(m, 2H), 3.74(m, 1H), 3.57(s, 2H), 3.20(s, 2H), 3.00(m, 4H ), 2.08(s, 1H).

Example 6 Preparation of His-Tyr-Trp (HYW)

1) Preparation of Boc-His(Boc)-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 433 mg (26%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 795 [M+H] ⁺ .

2) Preparation of His-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 430 mg (80%) of the title compound as a pale yellow powder. Mp: 118-120C; [α] _D ²⁵ =32.6 (c=1.04, CH ₃ OH); ESI ⁺ -MS (m/e) 505 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 11.01(s, 1H), 9.40(s, 1H), 8.99(d, J=8Hz, 1H), 8.93(s, 1H), 8.31(d, J=8.0Hz, 1H), 7.58(d , J=7.5Hz, 1H), 7.41(s, 1H), 7.32(d, J=7.5Hz, 1H), 7.16(s, 1H), 7.14(d, J=8.4Hz, 2H), 7.04(t , J=7.4Hz, 1H), 6.97(t, J=7.4Hz, 1H), 6.64(d, J=8.4Hz, 2H), 4.42(m, 2H), 3.30(m, 1H), 3.07(m , 1H), 2.95(m, 2H), 2.78(m, 2H), 2.08(s, 1H).

Embodiment 7 prepares Ile-Tyr-Trp (IYW)

1) Preparation of Boc-Ile-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 806 mg (84%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 691 [M+Na] ⁺ .

2) Preparation of Ile-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 290 mg (88%) of the title compound as a pale yellow powder. Mp: 168-169C; [α] _D ²⁵ =37.1 (c=0.97, CH ₃ OH); ESI ⁺ -MS (m/e) 481 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 10.88(s, 1H), 9.24(s, 1H), 8.53(d, J=8.1Hz, 1H), 8.43(d, J=7.5Hz, 1H), 7.53(d, J=7.5Hz, 1H), 7.34(d, J=7.5Hz, 1H), 7.17(s, 1H), 7.08(t, J=7.4Hz, 1H), 7.06(d, J=8.4Hz, 2H), 6.97(t, J=7.4Hz, 1H), 6.64(d, J=8.4Hz, 2H), 4.50(m, 2H), 3.17(m, 2H), 2.92(m, 2H), 2.68(m, 1H), 2.10( s, 1H), 1.09 (m, 2H), 0.89 (d, J = 7.0 Hz, 3H), 0.8 (t, J = 7.3 Hz, 3H).

Embodiment 8 prepares Lys-Tyr-Trp (KYW)

1) Preparation of Boc-Lys(Z)-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 640 mg (40%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 787 [M+H] ⁺ .

2) Preparation of Lys-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 280 mg (79%) of the title compound as a pale yellow powder. Mp: 131-132C; [α] _D ²⁵ =30.4 (c=1.24, CH ₃ OH); ESI ⁺ -MS (m/e) 496 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 12.68(s, 1H), 10.96(s, 1H), 9.25(s, 1H), 8.73(d, J=8.2Hz, 1H), 8.46(d, J=7.5Hz, 1H), 8.23( s, 2H), 8.03(s, 2H), 7.54(d, J=7.5Hz, 1H), 7.36(d, J=7.5Hz, 1H), 7.23(s, 1H), 7.09(t, J=7.4 Hz, 1H), 7.06(d, J=8.4Hz, 2H), 7.00(t, J=7.4Hz, 1H), 6.65(d, J=8.4Hz, 2H), 4.50(m, 2H), 3.45( m, 1H), 3.15(m, 2H), 2.90(m, 2H), 2.09(s, 3H), 1.75(m, 2H), 1.56(m, 2H), 1.47(m, 2H).

Example 9 Preparation of Leu-Tyr-Trp (LYW)

1) Preparation of Boc-Leu-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 1002 mg (83%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 671 [M+H] ⁺ .

2) Preparation of Leu-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 190 mg (50%) of the title compound as a pale yellow powder. Mp: 147-149C; [α] _D ²⁵ = 44.0 (c = 1.07, CH ₃ OH); ESI ⁺ -MS (m/e) 481.3 [M+H] ⁺ ; ¹ HNMR (BHSC-300Hz, DMSO-d ₆ ), 10.84(s, 1H), 9.20(s, 1H), 8.63(d, J=8Hz, 1H), 8.53(d, J=7.5Hz, 1H), 7.55(d, J=7.5Hz, 1H ), 7.38(d, J=7.5Hz, 1H), 7.17(s, 1H), 7.08(t, J=7.4Hz, 1H), 7.00(d, J=8.4Hz, 2H), 6.97(t, J =7.4Hz, 1H), 6.65(d, J=8.4Hz, 2H), 4.50(m, 2H), 3.45(m, 1H), 3.20(m, 4H), 2.09(s, 1H), 2.00(m , 1H), 1.64 (m, 2H).

Example 10 Preparation of Met-Tyr-Trp (MYW)

1) Preparation of Boc-Met-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 780 mg (80%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 711 [M+Na] ⁺ .

2) Preparation of Met-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 480 mg (80%) of the title compound as a pale yellow powder. Mp: 151-152C; [α] _D ²⁵ =20.7 (c=1.05, CH ₃ OH); ESI ⁺ -MS (m/e) 499 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 10.78(s, 1H), 9.21(s, 2H), 8.03(d, J=7.5Hz, 2H), 7.55(d, J=7.5Hz, 1H), 7.32(d, J=7.5Hz, 1H), 7.13(s, 1H), 7.04(t, J=7.4Hz, 1H), 6.97(m, 3H), 6.61(d, J=8.4Hz, 2H), 4.45(m, 1H), 4.35( m, 1H), 3.56(m, 1H), 3.23(m, 2H), 3.06(m, 2H), 2.39(t, J=6.7Hz, 2H), 2.05(m, 2H), 1.98(s, 3H ).

Embodiment 11 prepares Asn-Tyr-Trp (NYW)

1) Preparation of Boc-Asn-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 876 mg (78%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 672 [M+H] ⁺ .

2) Preparation of Asn-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 403 mg (82%) of the title compound as a pale yellow powder. Mp: 122-123C; [α] _D ²⁵ =22.4 (c=1.04, CH ₃ OH); ESI ⁺ -MS (m/e) 483 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 10.88(s, 1H), 9.44(s, 1H), 8.65(d, J=8.0Hz, 1H), 8.50(d, J=7.5Hz, 1H), 8.25(s, 2H), 7.60( d, J=7.5Hz, 1H), 7.33(d, J=7.5Hz, 1H), 7.20(s, 2H), 7.12(d, J=8.4Hz, 2H), 7.00(m, 3H), 6.69( d, J = 8.4 Hz, 2H), 4.50 (m, 2H), 3.00 (m, 6H), 2.09 (s, 1H).

Embodiment 12 prepares Pro-Tyr-Trp (PYW)

1) Preparation of Boc-Pro-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 1321 mg (92%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 655 [M+H] ⁺ .

2) Preparation of Pro-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 300 mg (82%) of the title compound as a pale yellow powder. Mp: 160-161C; [α] _D ²⁵ = -12.5 (c = 1.12, CH ₃ OH); ESI+-MS (m/e) 466 [M+H] ⁺ ; ¹ HNMR (BHSC-300Hz, DMSO-d ₆ ), 10.91(s, 1H), 9.21(s, 1H), 8.72(d, J=8.1Hz, 1H), 8.40(d, J=7.8Hz, 1H), 7.55(d, J=7.5Hz, 1H), 7.35(d, J=7.5Hz, 1H), 7.20(s, 1H), 7.03(d, J=8.4Hz, 2H), 6.99(m, 2H), 6.64(d, J=8.4Hz, 2H), 4.50(m, 2H), 4(m, 1H), 3.12(m, 4H), 2.95(m, 1H), 2.68(m, 1H), 2.09(s, 1H), 1.78(m, 4H ).

Example 13 Preparation of Gln-Tyr-Trp (QYW)

1) Preparation of Boc-Gln-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 730 mg (72%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 706 [M+Na] ⁺ .

2) Preparation of Gln-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 322 mg (83%) of the title compound as a pale yellow powder. Mp: 128-130C; [α] _D ²⁵ =31.5 (c=1.08, CH ₃ OH); ESI ⁺ -MS (m/e) 496 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 10.88(s, 1H), 9.18(s, 1H), 8.26(d, J=7.7Hz, 1H), 7.95(d, J=8.8Hz, 1H), 7.77(s, 1H), 7.55( d, J=7.5Hz, 1H), 7.33(d, J=7.5Hz, 1H), 7.17(s, 2H), 7.08(t, J=7.4Hz, 2H), 7.00(m, 3H), 6.58( d, J=8.4Hz, 2H), 4.50(m, 2H), 3.20(dd, J=5.0Hz, J=15.0Hz, 2H), 3.09(dd, J=7.8Hz, J=15.0Hz, 1H) , 2.92(dd, J=5.0Hz, J=15.0Hz, 1H), 2.68(dd, J=7.8Hz, J=15.0Hz, 1H), 2.02(m, 1H).

Embodiment 14 prepares Arg-Tyr-Trp (RYW)

1) Preparation of Boc-Arg(NO ₂ )-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 988 mg (86%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 759 [M+H] ⁺ .

2) Preparation of Arg-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 530 mg (50%) of the title compound as a pale yellow powder. Mp: 174-176C; [α] _D ²⁵ =45.2 (c=1.05, CH ₃ OH); ESI ⁺ -MS (m/e) 524 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 10.90(s, 1H), 9.28(s, 1H), 8.85(d, J=8.1Hz, 1H), 8.52(s, 2H), 8.04(d, J=7.5Hz, 1H), 7.60( d, J=7.5Hz, 1H), 7.37(s, 1H), 7.36(d, J=7.5Hz, 1H), 7.16(s, 1H), 7.06(m, 3H), 6.97(t, J=7.4 Hz, 1H), 6.62(d, J=8.4Hz, 2H), 4.35(m, 2H), 3.73(t, J=6.0Hz, 1H), 3.15(m, 2H), 2.90(m, 2H), 2.65(m, 2H), 1.91(s, 1H), 1.85(m, 2H), 1.55(m, 2H).

Example 15 Preparation of Ser-Tyr-Trp (SYW)

1) Preparation of Boc-Ser(-OBz1)-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 654 mg (34%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 645 [M+H] ⁺ .

2) Preparation of Ser-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 290 mg (85%) of the title compound as a yellow-free powder. Mp: 121-122C; [α] _D ²⁵ =9.8 (c=1.09, CH ₃ OH); ESI ⁺ -MS (m/e) 455 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 12.78(s, 1H), 10.91(s, 1H), 9.22(s, 1H), 8.60(d, J=8.1Hz, 1H), 8.42(d, J=7.5Hz, 1H), 8.13( s, 2H), 7.53(d, J=7.5Hz, 1H), 7.34(d, J=7.5Hz, 1H), 7.20(s, 1H), 7.05(d, J=8.4Hz, 1H), 6.98( m, 4H), 6.64(d, J=8.4Hz, 2H), 5.50(s, 1H), 4.50(m, 2H), 3.78(m, 2H), 3.09(m, 2H), 2.09(s, 1H ).

Embodiment 16 prepares Thr-Tyr-Trp (TYW)

1) Preparation of Boc-Thr-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 876 mg (87%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 651 [M+H] ⁺ .

2) Preparation of Thr-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 400 mg (90%) of the title compound as a non-yellow powder. Mp: 114-115C; [α] _D ²⁵ = 4.7 (c = 1.04, CH ₃ OH); ESI ⁺ -MS (m/e) 469 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 10.96(s, 1H), 8.66(d, J=8.1Hz, 1H), 8.52(d, J=7.5Hz, 1H), 8.18(s, 2H), 7.54(d, J=7.8Hz, 1H), 7.35(d, J=7.8Hz, 1H), 7.22(s, 1H), 7.06(m, 3H), 6.98(t, J=7.4Hz, 1H), 6.65(d, J=8.4Hz, 2H), 4.55(m, 3H), 3.91(m, 1H), 3.65(m, 1H), 3.21(dd, J=5.0Hz, J=15.0Hz, 1H), 3.08(dd, J=7.8Hz, J=15.0Hz, 1H), 2.95(dd, J=5.0Hz, J=15.0Hz, 1H), 2.72(dd, J=7.8Hz, J=15.0Hz, 1H), 2.10(s, 1H), 1.10 (d, J=6.3Hz, 3H).

Embodiment 17 prepares Val-Tyr-Trp (VYW)

1) Preparation of Boc-Val-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 789 mg (82%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 657 [M+H] ⁺ .

2) Preparation of Val-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 288 mg (89%) of the title compound as a yellow-free powder. Mp: 153-155C; [α] _D ²⁵ =7.3 (c=0.96, CH ₃ OH); ESI ⁺ -MS (m/e) 467 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 10.89(s, 1H), 9.24(s, 2H), 8.55(d, J=7.5Hz, 1H), 8.43(d, J=7.5Hz, 1H), 8.08(s, 2H), 7.53( d, J=7.5Hz, 1H), 7.33(d, J=7.5Hz, 1H), 7.16(s, 1H), 7.06(m, 3H), 6.97(t, J=7.4Hz, 1H), 6.61( d, J=8.4Hz, 2H), 4.55(m, 2H), 3.21(dd, J=5.5Hz, J=15.0Hz, 1H), 3.10(dd, J=7.8Hz, J=15.0Hz, 1H) , 2.92(dd, J=5.5Hz, J=15.0Hz, 1H), 2.75(dd, J=7.8Hz, J=15.0Hz, 1H), 2.13(m, 1H), 2.10(s, 1H), 1.07 (d, J = 7.0 Hz, 6H).

Embodiment 18 prepares Trp-Tyr-Trp (WYW)

1) Preparation of Boc-Trp-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 1098 mg (91%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 745 [M+H] ⁺ .

Boc-Trp-Tyr-Trp-OBz1(3s)

2) Preparation of Trp-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 340 mg (88%) of the title compound as a yellow-free powder. Mp: 171-172C; [α] _D ²⁵ =6.2 (c=1.03, CH ₃ OH); ESI ⁺ -MS (m/e) 554 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 11.00(s, 1H), 10.95(s, 1H), 9.24(s, 1H), 8.85(d, J=7.9Hz, 1H), 8.51(d, J=7.5Hz, 1H), 8.00( s, 2H), 7.57(d, J=7.5Hz, 2H), 7.35(d, J=7.5Hz, 2H), 7.24(s, 1H), 7.22(s, 1H), 7.08(d, J=8.4 Hz, 2H), 7.00(t, J=7.4Hz, 2H), 6.93(t, J=7.4Hz, 2H), 6.65(d, J=8.4Hz, 2H), 4.55(m, 2H), 4.00( s, 1H), 3.25(dd, J=5.0Hz, J=15.0Hz, 2H), 3.15(dd, J=7.8Hz, J=15.0Hz, 1H), 2.97(dd, J=5.0Hz, J= 15.0Hz, 2H), 2.75 (dd, J=7.8Hz, J=15.0Hz, 1H).

Example 19 Preparation of Tyr-Tyr-Trp (YYW)

1) Preparation of Boc-Tyr-Tyr-Trp-OBz1

Following the procedure for the preparation of Boc-Ala-Tyr-Trp-OBz1 afforded 694 mg (80%) of the title compound as a colorless solid. ESI ⁺ -MS (m/e) 722 [M+H] ⁺ .

2) Preparation of Tyr-Tyr-Trp

Following the procedure for the preparation of Ala-Tyr-Trp afforded 548 mg (88%) of the title compound as a yellow-free powder. Mp: 126-127C; [α] _D ²⁵ =10.5 (c=1.06, CH ₃ OH); ESI ⁺ -MS (m/e) 531 [M+H] ⁺ ; ¹ HNMR (BHSC-500Hz, DMSO-d ₆ ), 10.92(s, 1H), 9.35(s, 1H), 9.30(s, 1H), 8.78(d, J=8.0Hz, 1H), 8.48(d, J=8.5Hz, 1H), 7.97( s, 2H), 7.57(d, J=7.5Hz, 2H), 7.35(d, J=7.5Hz, 2H), 7.21(s, 1H), 7.06(m, 6H), 6.99(t, J=7.4 Hz, 2H), 6.69(d, J=8.4Hz, 2H), 6.64(d, J=8.4Hz, 2H), 4.55(m, 2H), 3.15(dd, J=5.0Hz, J=15.0Hz, 1H), 3.00(dd, J=7.8Hz, J=15.0Hz, 2H), 2.92(dd, J=5.0Hz, J=15.0Hz, 1H), 2.68(dd, J=7.8Hz, J=15.0Hz , 2H).

Experimental example 1 The analgesic effect evaluation experiment of the compound of the present invention

1. Test compounds: 19 compounds prepared in Examples 1-19 of the present invention;

2. Experimental method:

Male ICR mice with a body weight of (22±2g) were exposed to light for 12 hours a day, the indoor temperature was 20-24°C, the humidity was maintained at 45-65%, and sufficient food and water were provided. The day before the experiment began to fast and feed, and the mice were put into the experimental operation room to adapt to the environment, and each mouse only participated in the experiment once.

During the experiment, the mice were put into a special fixed cylinder, the tail was exposed to the outside, and 75% alcohol was used for disinfection. When the pain threshold was measured, the light beam generated by a special spotlight was used to irradiate the lower third of the mouse tail, and a stopwatch was used for timing. The latency from irradiation to tail flick response (tail flick latency, TFL) was used as the pain threshold. Adjust the distance between the spotlight and the mouse tail to make the stimulus intensity more sensitive and the TFL to be 2-6s. At the beginning of the experiment, three times were measured at an interval of 5 minutes each time, and the mean value was taken as the basic pain threshold. The test compounds were respectively suspended in 0.5% carboxymethylcellulose sodium (CMC-Na) aqueous solution, administered by intragastric administration, and the suspension concentration was adjusted according to the dosage of 0.2ml and 0.13mmol/kg for each intragastric administration. Each experiment set up 0.5% CMC-Na group as parallel control. In order to prevent skin burns, the light cut-off time was 10s, and the TFL was retested at intervals of 30 minutes to observe the relationship between the time and effect of drug analgesia. The intensity of drug analgesia was evaluated by the increase rate of pain threshold: increase rate of pain threshold = (post-administration pain threshold - basic pain threshold) ÷ basic pain threshold. The obtained results are listed in Table 1, and the results show that the compound of the present invention has definite analgesic effect.

Table 1 The analgesic activity test result of compound of the present invention

Note: the increase rate of pain threshold is represented by mean ± SD%, n=10, and the dosage is 0.13mmol/kg;

a) Compared with CMC-Na group, P<0.05;

b) Compared with CMC-Na group, P<0.01;

c) P<0.001 compared with CMC-Na group.

Analgesic activity of experimental example 2 different doses of AYW and KYW

1) Test compound: compounds AYW and KYW prepared in the examples of the present invention;

2) Experimental method: The analgesic effect of the test compound at three doses of 0.13mmol/kg, 0.08mmol/kg and 0.05mmol/kg was evaluated using the light radiation heat tail flick model of mice in Experimental Example 1. The results are listed in Table 2. The experimental results showed that all the tested compounds exhibited analgesic effect in a dose-dependent manner.

Table 2 Analgesic effects of different doses of AYW and KYW

Note: The increase rate of pain threshold is represented by X±SD%, n=10; a) P<0.05 compared with the 0.08mmol/kg group; b) P<0.05 compared with the 0.05mmol/kg group.

Claims (9)

1. The aminoacyl tyrosyl tryptophan tripeptide with analgesic activity, its structural formula is shown in general formula I: AA-Tyr-Trp I Wherein, AA is an alanine residue. 2. A method for preparing the aminoacyltyrosyltryptophan tripeptide with analgesic activity according to claim 1, comprising: (1) coupling tert-butoxycarbonyl-protected tyrosine and tryptophan benzyl ester to synthesize tert-butoxycarbonyl tyrosyl tryptophan benzyl ester; (2) removing the tert-butoxycarbonyl group of tert-butoxycarbonyl tyrosyl tryptophan benzyl ester to obtain tyrosyl tryptophan benzyl ester; (3) Coupling tyrosyl tryptophan benzyl ester with tert-butoxycarbonyl-protected alanine to obtain tert-butoxycarbonylalanyl tyrosyl tryptophan benzyl ester; (4) hydrogenolysis and debenzylation of benzyl tert-butoxycarbonylalanyl tyrosyl tryptophanate to obtain tert-butoxycarbonyl alanyl tyrosyl tryptophan; (5) Remove tert-butoxycarbonyl from tert-butoxycarbonylalanyl tyrosyl tryptophan to obtain the aminoacyl tyrosyl tryptophan tripeptide of general formula I. 3. according to the described method of claim 2, it is characterized in that: in the step (1) in the presence of dicyclohexylcarbodiimide, N-hydroxybenzotriazole, N-methylmorpholine and anhydrous THF tert-butoxycarbonyl-protected tyrosine is coupled with tryptophan benzyl ester to synthesize tert-butoxycarbonyl tyrosyl tryptophan benzyl ester. 4. according to the method for claim 2, it is characterized in that: in step (2) in hydrogen chloride/ethyl acetate solution, add tert-butoxycarbonyl tyrosyl tryptophan benzyl ester, slough tert-butoxycarbonyl, prepare Tyrosyl tryptophan benzyl ester is obtained. 5. according to the described method of claim 2, it is characterized in that: in the step (3) in the presence of dicyclohexylcarbodiimide, N-hydroxybenzotriazole, N-methylmorpholine and anhydrous THF Tyrosyl tryptophan benzyl ester is coupled with tert-butoxycarbonyl-protected alanine to obtain tert-butoxycarbonylalanyl tyrosyl tryptophan benzyl ester. 6. according to the method for claim 2, it is characterized in that: in the step (4) in the presence of palladium carbon, tert-butoxycarbonylalanyl tyrosyl tryptophan benzyl ester hydrogenolysis debenzylation, prepare tert-butyl Oxycarbonylalanyl tyrosyl tryptophan. 7. The method according to claim 2, characterized in that: in step (5), t-butoxycarbonyl alanyl tyrosyl tryptophan is removed from tert-butoxycarbonyl in hydrogen chloride/ethyl acetate solution. 8. A pharmaceutical composition for treating pain, comprising an effective amount of the aminoacyltyrosyltryptophan tripeptide according to claim 1 and a pharmaceutically acceptable carrier or auxiliary material. 9. The use of the aminoacyltyrosyltryptophan tripeptide according to claim 1 in the preparation of analgesic drugs.

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