JPH0586096A - Peptide derivative of dideoxynucleoside and anti-HIV agent containing the same - Google Patents

Abstract

(57) [Summary] (Modified) [Objective] A peptide derivative of dideoxynucleoside is synthesized and used as an anti-HIV agent. [Structure] A peptide derivative of a dideoxynucleoside represented by the formula [1] or [2], and an anti-HIV agent containing the peptide derivative of the dideoxynucleoside as an active ingredient. In addition, these peptide derivatives are prepared by reacting dideoxynucleoside with oxalyl dichloride or oxalyl chloride monoester and then producing amino acids 2 to 1
It is synthesized by reacting with 0 peptide ester. [Wherein R ¹ is a hydrogen atom, R ² is an alkyl group, a hydroxymethyl group, R ³ is a benzyl group which may have a substituent, X is a lower alkoxy, benzyloxy or an amino acid or peptide derivative, Y Is a hydrogen atom or N ₃ , B
Represents a nucleic acid base]

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a peptide derivative of dideoxynucleoside and an anti-human immunodeficiency virus agent containing the same as an active ingredient.

[0002]

2. Description of the Related Art Acquired immunodeficiency syndrome [Acquire]
d Immunity Deficiency Syndr
ome; hereinafter abbreviated as AIDS] is an anti-human immunodeficiency virus [Human Immunodeficiency
Virus; hereinafter abbreviated as HIV], is a serious immunodeficiency disease caused by an extremely high mortality rate, and measures against such HIV infection and AIDS have become a major social issue. Currently, as an anti-HIV agent which is recognized to be clinically effective, azidothymidine [hereinafter abbreviated as AZT] having a reverse transcriptase inhibitory action is known, but its clinical effect is still insufficient. Further, there is a problem that side effects due to this, for example, side effects such as injury of bone marrow (hematopoietic tissue), headache, and neurological symptoms such as convulsions are strong. In particular, since HIV has a gene that sneak into a condition like a genetic disease, long-term administration of a drug is inevitably required, and the side effect of AZT is a serious injury when it is used as an anti-HIV agent. Has become.

[0003]

[Problems to be Solved by the Invention] HIV infected person has AID
There is usually an extremely long clinical incubation period before the onset of S, which makes it very difficult to take preventive measures against infection. Under such circumstances, development of new pharmaceutical preparations that are effective against HIV infection and AIDS has been awaited.

[0004]

Means for Solving the Problems As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that a reverse transcriptase inhibitor such as AZT is a peptide having a binding active site peptide of CD4 to gp120, such as L-. Ceryl-L-phenylalanyl-
L-leucyl-L-threonyl-L-lysyl-glycyl-L-prolyl-L-serine (Ser-Phe-Leu
-Thr-Lys-Gly-Pro-Ser-OH),
D- prolyl -L- phenylalanine benzyl ester (D-Pro-L-Phe -OCH 2 Ph) and L- prolyl -L- phenylalanine benzyl ester (L-
The present invention was completed by finding that a compound obtained by binding Pro-L-Phe-OCH2Ph) with an oxalyl group has an excellent anti-HIV activity and high safety.

That is, the general formula [1]

[0006]

[Chemical 5]

(In the formula, R ₁ is a hydrogen atom, R ₂ is an alkyl group, a hydroxymethyl group, or — (CH) _n — in which R ₁ and R ₂ are connected, n is 3 or 4, and R ³ is A benzyl group which may have a substituent, X is a lower alkoxy,
A benzyloxy group, an amino acid or a peptide derivative, Y represents a hydrogen atom or an azido group, and B represents a nucleobase), and a peptide derivative of a dideoxynucleoside represented by the formula and an anti-HIV containing the same as an active ingredient
To provide the agent.

In the general formula [1], R ¹ is a hydrogen atom, R ² is an alkyl group, a hydroxymethyl group, or R ¹
And R ² are connected to each other,-(CH) _n -in which n is 3 or 4, and may form a 5-membered ring or a 6-membered ring. The asymmetric carbon atom to which R ² is bonded may be the (L) body or the (D) body.
When R ¹ is a hydrogen atom, R ² is a usual amino acid residue, preferably a hydroxymethyl group corresponding to serine. R ³ is a benzyl group which may have a substituent, that is, an aromatic amino acid residue, but a benzyl group corresponding to phenylalanine is preferable. X is lower alkoxy, benzyloxy group, amino acid or peptide derivative,
Y represents a hydrogen atom or an azide group, and B represents a nucleic acid base.

The peptide derivative [1] of the dideoxynucleoside of the present invention is prepared by reacting a nucleobase having an azido group or a hydrogen atom at the 3'position with oxalyl chloride or p-nitrophenyl oxalyl chloride in the presence of a base, It can be obtained by reacting with a peptide ester and removing the ester group or peptide protecting group if necessary. Examples of the nucleobase include 3'-azido-3'-deoxythymidine [abbreviated as AZT], 2 ', 3'-dideoxyuridine [abbreviated as ddU], 2', 3'-dideoxycytidine [abbreviated as ddC]. 2 ′, 3′-dideoxyinosine [abbreviated as ddI] and 2 ′, 3′-dideoxyadenosine [abbreviated as ddA]. Examples of the base include triethylamine, and 4-dimethylaminopyridine is also used if necessary.
The peptide ester is benzyl or C1-4 of a peptide consisting of 2 to 10 amino acids of (L) or (D) form.
Is a lower alkyl ester consisting of

The peptide derivative of the dideoxynucleoside represented by the general formula [1] of the present invention is used as an essential component, and usually it is prepared as a drug by adding a suitable pharmaceutical preparation carrier together with its pharmacologically effective amount. As a pharmaceutical carrier,
Any of fillers, extenders, moisturizers, disintegrants, excipients such as surfactants and diluents or the like which are commonly used for preparing a preparation depending on the form of use can be used. The form of the pharmaceutical composition is not particularly limited as long as it effectively contains the above-mentioned active ingredient, and examples thereof include solid agents such as tablets, powders, granules and pills, and ordinary liquids, suspensions, emulsions and the like. It can be a liquid. Further, it can be made into a dried product which can be made into a liquid form by adding an appropriate carrier before use. If necessary, various usual additives such as solubilizers, buffers, soothing agents, preservatives, coloring agents, etc. may be added to the pharmaceutical composition, and further other pharmaceuticals may be added in combination. You can also The anti-HIV agent of the present invention is administered by an appropriate administration route depending on the form of the pharmaceutical composition. The administration method is not particularly limited, and may be internal use, external use and injection.

The dose of the drug containing the peptide derivative of the dideoxynucleoside represented by the general formula [1] as an active ingredient as an anti-HIV agent depends on its dosage form, administration method, intended use and age of the patient to whom it is applied. The amount of the active ingredient contained in the preparation is generally 0 in the case of oral administration per adult, although it is appropriately set depending on the body weight, medical condition, etc.
It is preferably about 1 g to 10 g, and about 0.1 g to 5 g for parenteral administration. In addition, administration is 1
It is also possible to divide into several times a day.

[0012]

INDUSTRIAL APPLICABILITY The peptide derivative [1] of dideoxynucleoside of the present invention is effective for the prevention of HIV infection and the prevention and treatment of AIDS, and its low cytotoxicity makes it suitable for long-term administration. It can be used.
Therefore, the anti-HI of the present invention containing this as an active ingredient
V agent is extremely useful for the prevention and treatment of such HIV infection in healthy people and the treatment of AIDS patients who have developed it.

[0013]

The present invention will be described in more detail with reference to Reference Examples, Examples and Test Examples, but the present invention is not limited to these.

[0014]

Reference Example 1 Oxalyl chloride (6 ml) was added to p-nitrophenol (750 mg), and the mixture was heated under reflux and boiled for 16 hours in an argon atmosphere. The reaction mixture was concentrated under reduced pressure, ether was added, and the mixture was filtered. The filtrate was concentrated to dryness to obtain p-nitrophenyl oxalyl chloride (350 mg). Melting point 85-90 ° C. IR (KBr) ν: 1785,170
6.

[0015]

[Reference Example 2] N-tert-butoxycarbonyl-L-
Proline (215 mg) and L-phenylalanine benzyl ester p-toluenesulfonate (428 m
g) was dissolved in N, N-dimethylformamide [hereinafter abbreviated as DMF] (5 ml) under an argon atmosphere, and diethyl phosphonocyanidate [hereinafter abbreviated as DEPC] under ice cooling.
(179 mg) was added. Then triethylamine (2
(12 mg) was added dropwise, and the mixture was stirred under ice cooling for 4 hours and further at room temperature for 18 hours. After concentration under reduced pressure, methylene chloride (1
(00 ml) was added, and the mixture was washed successively with aqueous sodium hydrogen carbonate, 10% aqueous citric acid, water and brine, dried over anhydrous magnesium sulfate and concentrated. This is subjected to silica gel chromatography (n-hexa
ne / AcOEt = 2/1), N-ter
t-Butoxycarbonyl-L-prolyl-L-phenylalanine benzyl ester (420 mg) was obtained as a colorless oil. [Α] _D −52.8 ° (c1.41, CHCl ₃ , 22
° C). FABMS (m / z): 453 (M + H) ⁺ . I
R (KBr) ν: 3450 (NH), 1745 (est
er), 1632, 1540 (amide). ¹ H-N
MR (CDCl ₃ ) δ: 7.38 to 7.05 (10H,
m, Ar), 5.15 (2H, br s, COOC H ₂
Ph), 4.28, 4.18 (1H, each br)
s, Pro; NCHCO), 4.14-4.09 (1
H, m, Phe; NHC H CO), 3.43~3.27
(2H, m, Pro; C H ₂ NCH), 3.18, 3.
02 (2H, m, Phe; CHCH ₂ Phe), 2.6.
_{2~1.70 (4H, m, Pro;} C H 2 C H 2 CHC
O), 1.41 (9H, br s, OC (C
H _{3) 3).}

[0016]

[Reference Example 3] N-tert-butoxycarbonyl-L-
Prolyl-L-phenylalanine benzyl ester (8
5 mg) was dissolved in a 4N-hydrogen chloride-ethyl acetate solution. Stir for 30 minutes at room temperature. After concentration under reduced pressure and dissolution in methylene chloride (10 ml), Amberlite IRA 94
0 (700 mg) was added, and the mixture was stirred at room temperature for 20 minutes. The insoluble material was filtered off, and concentrated under reduced pressure to give L-prolyl-L-phenylalanine benzyl ester (65 mg) as a colorless oil.

[0017]

[Reference Example 4] N-tert-butoxycarbonyl-L-
Proline (215 mg) and D-phenylalanine benzyl ester p-toluenesulfonate (428 m
g) was dissolved in DMF (5 ml) under an argon atmosphere, and DEPC (179 mg) was added under ice cooling. Then, triethylamine (212 mg) was added dropwise, and the mixture was stirred under ice cooling for 4 hours, and further stirred at room temperature for 18 hours. After concentration under reduced pressure, methylene chloride (100 ml) was added, and the mixture was washed successively with aqueous sodium hydrogen carbonate, 10% aqueous citric acid, water and brine, dried over anhydrous magnesium sulfate and concentrated. This was purified by silica gel chromatography (n-hexane / AcOEt = 2/1) and N-tert-butoxycarbonyl-L-prolyl-D-phenylalanine benzyl ester (420 m
g) was obtained as an oil. [Α] _D −45.7 ° (c
1.22, CHCl ₃ , 22 ° C.). FABMS (m /
z): 453 (M + H) ⁺ . IR (KBr) ν: 344
0 (NH), 1746 (ester), 1635, 15
37 (amide). ¹ H-NMR (CDCl ₃ ) δ:
7.38 to 7.05 (10H, m, Ar), 5.15
(2H, br s, COOC H ₂ Ph), 4.28,
4.18 (1H, each br s, Pro; NCH
CO), 4.14-4.09 (1H, m, Phe; NH
C H CO), 3.43~3.27 (2H , m, Pro;
CH ₂ NCH), 3.18, 3.02 (2H, m, Ph
e; CHCH ₂ Phe), 2.62-1.70 (4H,
_{m, Pro; C H 2 C} H 2 CHCO), 1.41 (9
H, br s, OC (C H ₃ ) ₃ ).

[0018]

[Reference Example 5] N-tert-butoxycarbonyl-L-
Prolyl-D-phenylalanine benzyl ester (8
5 mg) was dissolved in a 4N-hydrogen chloride-ethyl acetate solution. Stir for 30 minutes at room temperature. After concentration under reduced pressure and dissolution in methylene chloride (10 ml), Amberlite IRA 94
0 (700 mg) was added, and the mixture was stirred at room temperature for 20 minutes. The insoluble material was filtered off, and concentrated under reduced pressure to give L-prolyl-D-phenylalanine benzyl ester (65 mg) as a colorless oil.

[0019]

[Reference Example 6] N-carbobenzoxy-L-serine (3.
0 g) was suspended in methylene chloride (150 ml), concentrated sulfuric acid (4 drops) was added, and then isobutylene was bubbled in for 20 minutes. After stirring at room temperature for 3 hours, the mixture was neutralized and washed with aqueous sodium hydrogen carbonate, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. This is subjected to silica gel chromatography (n-hexane / AcOEt).
= 10/1), N-carbobenzoxy-O
-Tert-Butyl-L-serine tert-butyl ester (2.83 g) was obtained as an oil. [Α]
_D 8.5 ° (c1.72, CHCl ₃ ). FABMS
(M / z): 352 (M + H) ⁺ . IR (KBr) ν:
3450 (NH), 1740 (ester), 168
0,1510 (amide).

[0020]

Reference Example 7 N-carbobenzoxy-O-tert-butyl-L-serine tert-butyl ester (2.8
3 g) was dissolved in methanol (150 ml), 5% -palladium-carbon (50% water content, 1.0 g) was added, and the mixture was stirred under a hydrogen atmosphere for 6 hours and catalytic reduction was performed. After removing the catalyst by filtration, the mixture was concentrated under reduced pressure at room temperature to obtain O-tert-butyl-L-serine tert-butyl ester (1.74 g).

[0021]

[Reference Example 8] O-tert-butyl-L-serine te
rt-Butyl ester (1.74 g) and N-carbobenzoxy-L-proline (2.0 g) were dissolved in DMF (10 ml) under an argon atmosphere, and DEPC (1.4 was prepared under ice cooling).
4 g) was added, triethylamine (891 mg) was added dropwise, and the mixture was stirred at room temperature for 18 hours. After concentration under reduced pressure, methylene chloride (250 ml) was added, and sodium bicarbonate water, 10% citric acid water,
The extract was washed successively with water and brine, dried over anhydrous magnesium sulfate and concentrated. Silica gel chromatography (n
-Hexane / AcOEt = 2/1),
N-carbobenzoxy-L-prolyl-O-tert-
Butyl-L-serine tert-butyl ester (3.
11 g) was obtained as an oil. [Α] _D −29.7 °
(C0.97, CHCl _3). FABMS (m / z):
449 (M + H) ⁺ . IR (KBr) ν: 3340 (N
H), 1735 (ester), 1650, 1525.
(Amide).

[0022]

[Reference Example 9] N-carbobenzoxy-L-prolyl-O
-Tert-Butyl-L-serine tert-butyl ester (3.11 g) was dissolved in methanol (150 ml), and 5% -palladium-carbon (containing 50% water, 1.0 g).
Was added, and the mixture was stirred under a hydrogen atmosphere for 6 hours and catalytic reduction was performed.
After removing the catalyst by filtration, the mixture was concentrated under reduced pressure at room temperature and then L-prolyl-Ot
ert-Butyl-L-serine tert-butyl ester (2.15 g) was obtained.

[0023]

[Reference Example 10] L-prolyl-O-tert-butyl-
L-serine tert-butyl ester (2.15 g)
And N-carbobenzoxyglycine (1.43 g) were dissolved in DMF (25 ml) under an argon atmosphere, and DE was added under ice cooling.
PC (1.23 g) was added, and triethylamine (762
(mg) was added dropwise, and the mixture was stirred for 4 hours and then at room temperature for 18 hours. After concentration under reduced pressure, methylene chloride (250 ml) was added, and the mixture was washed successively with aqueous sodium hydrogen carbonate, 10% citric acid water, water and brine, dried over anhydrous magnesium sulfate and concentrated. This was subjected to silica gel chromatography (n-hexane / AcOEt =
1/10) to give N-carbobenzoxyglycyl-L-prolyl-O-tert-butyl-L-serine tert-butyl ester (3.35 g) as an oil. [Α] _D −43.7 ° (c1.08, CHC
l ₃ ). FABMS (m / z): 506 (M + H) ⁺ .
IR (KBr) ν: 3320 (NH), 1735 (es)
ter), 1659, 1525 (amide).

[0024]

[Reference Example 11] N-carbobenzoxyglycyl-L-prolyl-O-tert-butyl-L-serine tert
-Butyl ester (3.35 g) in methanol (150
ml), 5% -palladium-carbon (50% water content, 1.0 g) was added, and the mixture was stirred under a hydrogen atmosphere for 6 hours and catalytic reduction was performed. After removing the catalyst by filtration, the mixture was concentrated under reduced pressure at room temperature to obtain glycyl-L-prolyl-O-tert-butyl-L-serine tert-butyl ester (2.41 g).

[0025]

Reference Example 12 Glycyl-L-prolyl-O-tert
-Butyl-L-serine tert-butyl ester (2.41 g) and N (α) -carbobenzoxy-N
(Ε) -tert-Butoxycarbonyl-L-lysine (2.47 g) under argon atmosphere in DMF (20 ml).
After dissolution, DEPC (1.17 g) was added under ice cooling, triethylamine (722 mg) was added dropwise, and the mixture was stirred for 4 hours and then at room temperature for 18 hours. After concentration under reduced pressure, methylene chloride (2
(50 ml) was added, and the mixture was washed successively with aqueous sodium hydrogen carbonate, 10% aqueous citric acid, water and brine, dried over anhydrous magnesium sulfate and concentrated. This is subjected to silica gel chromatography (CHCl ₃ /
MeOH = 30/1) and purified by N (α) -carbobenzoxy-N (ε) -tert-butoxycarbonyl-L-lysyl-glycyl-L-prolyl-O-tert.
-Butyl-L-serine tert-butyl ester (3.0 g) was obtained. _{[Α] D -35.3 ° (c1.39} , CHCl 3). F
ABMS (m / z): 734 (M + H) ⁺ . IR (KB
r) ν: 3342 (NH), 1733 (ester),
1663, 1525 (amide).

[0026]

[Reference Example 13] N (α) -carbobenzoxy-N (ε)
-Tert-Butoxycarbonyl-L-lysyl-glycyl-L-prolyl-O-tert-butyl-L-serine tert-butyl ester (3.00 g) was dissolved in methanol (150 ml), and 5% -palladium-carbon ( 50% water content, 1.0 g) was added, and the mixture was stirred and catalytically reduced in a hydrogen atmosphere for 6 hours. After removing the catalyst by filtration, the mixture was concentrated under reduced pressure at room temperature and N (ε) -tert-butoxycarbonyl-L-lysyl-glycyl-L-prolyl-O-tert-butyl-L-serine tert-butyl ester (2.43) was added.
g) was obtained.

[0027]

Reference Example 14 N (ε) -tert-butoxycarbonyl-L-lysyl-glycyl-L-prolyl-O-ter
t-Butyl-L-serine tert-butyl ester (2.43 g) and N-carbobenzoxy-O-ter.
t-Butyl-L-threonine (1.25 g) was dissolved in DMF (20 ml) under an argon atmosphere, and DEPC was performed under ice cooling.
(728 mg) was added, and triethylamine (451 m
After g) was added dropwise, the mixture was stirred for 4 hours and then at room temperature for 18 hours. After concentration under reduced pressure, methylene chloride (250 ml) was added, and the mixture was washed successively with aqueous sodium hydrogen carbonate, 10% citric acid water, water and brine, dried over anhydrous magnesium sulfate and concentrated. This was subjected to silica gel chromatography (CHCl ₃ / MeOH = 30 /
Purified by 1), N-carbobenzoxy-O-ter
t-Butyl-L-threonyl-N (ε) -tert-butoxycarbonyl-L-lysyl-glycyl-L-prolyl-O-tert-butyl-L-serine tert-butyl ester (3.06 g) was obtained. Melting point 66-73
° C. [Α] _D -15.5 ° (c0.99, CHC
l ₃ ). FABMS (m / z): 892 (M + H) ⁺ .
IR (KBr) ν: 3400 (NH), 1735 (es)
ter), 1642, 1525 (amide).

[0028]

[Reference Example 15] N-carbobenzoxy-O-tert-
Butyl-L-threonyl-N (ε) -tert-butoxycarbonyl-L-lysyl-glycyl-L-prolyl-
O-tert-butyl-L-serine tert-butyl ester (3.06 g) was dissolved in methanol (150 ml), and 5% -palladium-carbon (50% water content, 1.0).
g) was added, and the mixture was stirred in a hydrogen atmosphere for 6 hours and catalytically reduced. After removing the catalyst by filtration, the mixture was concentrated under reduced pressure at room temperature to give O-tert-butyl-L-threonyl-N (ε) -tert-butoxycarbonyl-L-lysyl-glycyl-L-prolyl-O.
-Tert-Butyl-L-serine tert-butyl ester (2.57 g) was obtained.

[0029]

Reference Example 16 O-tert-butyl-L-threonyl-N (ε) -tert-butoxycarbonyl-L-lysyl-glycyl-L-prolyl-O-tert-butyl-
L-serine tert-butyl ester (2.57 g)
And N-carbobenzoxy-L-leucine (0.90
g) was dissolved in DMF (20 ml) under an argon atmosphere, DEPC (611 mg) was added under ice cooling, triethylamine (378 mg) was added dropwise, and the mixture was stirred for 4 hours and then at room temperature for 18 hours. After concentration under reduced pressure, methylene chloride (250 ml)
Was added, and the mixture was washed successively with aqueous sodium hydrogen carbonate, 10% aqueous citric acid, water and brine, dried over anhydrous magnesium sulfate and concentrated. This was subjected to silica gel chromatography (CHCl ₃ / MeOH =
30/1) and N-carbobenzoxy-L-
Leucyl-O-tert-butyl-L-threonyl-N
(Ε) -tert-Butoxycarbonyl-L-lysyl-
Glycyl-L-prolyl-O-tert-butyl-L-
Serine tert-butyl ester (2.95 g) was obtained. Melting point 94-96 ° C. [Α] _D −26.9 ° (c1.
04, CHCl _3). FABMS (m / z): 1005
(M + H) ⁺ . IR (KBr) ν: 3320 (NH),
1737 (ester), 1653, 1520 (ami
de).

[0030]

Reference Example 17 N-carbobenzoxy-L-leucyl-
O-tert-butyl-L-threonyl-N (ε) -t
ert-butoxycarbonyl-L-lysyl-glycyl-
L-prolyl-O-tert-butyl-L-serine t
ert-Butyl ester (2.95 g) was dissolved in methanol (150 ml) and 5% -palladium-carbon (5
0% water content, 1.0 g) was added, and the mixture was stirred and catalytically reduced in a hydrogen atmosphere for 6 hours. After removing the catalyst by filtration, the mixture was concentrated under reduced pressure at room temperature and then L-leucyl-O-tert-butyl-L-threonyl-N (ε) -tert-butoxycarbonyl-L-lysyl-glycyl-L-prolyl-O-tert-butyl. -L-serine tert-butyl ester (2.50
g) was obtained.

[0031]

[Reference Example 18] L-leucyl-O-tert-butyl-
L-threonyl-N (ε) -tert-butoxycarbonyl-L-lysyl-glycyl-L-prolyl-O-te
rt-Butyl-L-serine tert-butyl ester (2.50 g) and N-carbobenzoxy-L-phenylalanine (0.811 g) under argon atmosphere in DMF.
(20 ml) was dissolved, DEPC (528 mg) was added under ice-cooling, triethylamine (378 mg) was added dropwise, and the mixture was stirred for 4 hours and then at room temperature for 18 hours. After concentration under reduced pressure, saturated aqueous sodium hydrogen carbonate was added and the precipitated crystals were collected by filtration. This was subjected to silica gel chromatography (CHCl ₃ / MeOH = 25 /
Purified by 1), N-carbobenzoxy-L-phenylalanyl-L-leucyl-O-tert-butyl-L
-Threonyl-N (ε) -tert-butoxycarbonyl-L-lysyl-glycyl-L-prolyl-O-ter
t-Butyl-L-serine tert-butyl ester (2.85 g) was obtained. Melting point 182-185 [deg.] C. [Α] _D −37.81 ° (c1.18, CHCl ₃ / M
eOH = 1/1). FABMS (m / z): 1152
(M + H) ⁺ . IR (KBr) ν: 3300 (NH),
1734 (ester), 1642, 1540 (ami
de).

[0032]

Reference Example 19 N-carbobenzoxy-L-phenylalanyl-L-leucyl-O-tert-butyl-L-threonyl-N (ε) -tert-butoxycarbonyl-
L-lysyl-glycyl-L-prolyl-O-tert-
Butyl-L-serine tert-butyl ester (2.
85 g) was dissolved in methanol (200 ml), and 5%-
Palladium-carbon (containing 50% water, 1.0 g) was added, and the mixture was stirred under a hydrogen atmosphere for 12 hours and catalytically reduced. After removing the catalyst by filtration, the mixture was concentrated under reduced pressure at room temperature and then L-phenylalanyl-L-leucyl-O-tert-butyl-L-threonyl-N.
(Ε) -tert-Butoxycarbonyl-L-lysyl-
Glycyl-L-prolyl-O-tert-butyl-L-
Serine tert-butyl ester (2.49 g) was obtained.

[0033]

[Reference Example 20] L-phenylalanyl-L-leucyl-
O-tert-butyl-L-threonyl-N (ε) -t
ert-butoxycarbonyl-L-lysyl-glycyl-
L-prolyl-O-tert-butyl-L-serine t
ert-Butyl ester (2.49 g) and N-carbobenzoxy-O-tert-butyl-L-serine (0.
724 g) was dissolved in DMF (20 ml) under an argon atmosphere, DEPC (440 mg) was added under ice-cooling, triethylamine (273 mg) was added dropwise, and the mixture was stirred for 4 hours and then at room temperature for 18 hours. After concentration under reduced pressure, methylene chloride (250
ml) was added and the mixture was washed successively with aqueous sodium hydrogen carbonate, 10% aqueous citric acid, water and brine, dried over anhydrous magnesium sulfate and concentrated.
This is subjected to silica gel chromatography (CHCl ₃ / Me)
OH = 25/1) and N-carbobenzoxy-O-tert-butyl-L-seryl-L-phenylalanyl-L-leucyl-O-tert-butyl-L-threonyl-N (ε). -Tert-butoxycarbonyl-
L-lysyl-glycyl-L-prolyl-O-tert-
Butyl-L-serine tert-butyl ester (2.
41 g) was obtained. Melting point 198-201 ° C. [Α] _D -2
6.6 ° (c1.07, CHCl ₃ ). FABMS (m
/ Z): 1295 (M + H) ⁺ . IR (KBr) ν: 3
400 (NH), 1735 (ester), 1634,
1521 (amide). Elemental analysis: Found. C6
0.95; H8.26; N9.67. Calcd. C6
0.89; H8.39; N9.54.

[0034]

[Reference Example 21] N-carbobenzoxy-O-tert-
Butyl-L-seryl-L-phenylalanyl-L-leucyl-O-tert-butyl-L-threonyl-N
(Ε) -tert-Butoxycarbonyl-L-lysyl-
Glycyl-L-prolyl-O-tert-butyl-L-
Serine tert-butyl ester (500 mg) was dissolved in methanol (150 ml), and 5% -palladium-
Carbon (50% water content, 0.3g) was added and under hydrogen atmosphere 1
The mixture was stirred for 2 hours and catalytically reduced. After removing the catalyst by filtration, the mixture was concentrated under reduced pressure at room temperature and O-tert-butyl-L-seryl-L-phenylalanyl-L-leucyl-O-tert-butyl-L-threonyl-N (ε) -tert-butoxycarbonyl was added. -L-lysyl-glycyl-L-prolyl-Ot
ert-Butyl-L-serine tert-butyl ester (444 mg) was obtained.

[0035]

[Reference Example 22] AZT (50 mg) in THF (1 ml)
Dissolved in O-tert-butyl-L-seryl-
L-phenylalanyl-L-leucyl-O-tert-
Butyl-L-threonyl-N (ε) -tert-butoxycarbonyl-L-lysyl-glycyl-L-prolyl-
O-tert-butyl-L-serine tert-butyl ester (326 mg) and DMAP (12 mg) were added, and the mixture was stirred at room temperature under an argon atmosphere for 24 hours. After concentration under reduced pressure, silica gel chromatography (CHCl ₃ / Me
OH = 8/1), N-carbo- (3′-azido-3′-deoxythymidyloxycarbonyl) -O
-Tert-butyl-L-seryl-L-phenylalanyl-L-leucyl-O-tert-butyl-L-threonyl-N (ε) -tert-butoxycarbonyl-L-
Lysyl-glycyl-L-prolyl-O-tert-butyl-L-serine tert-butyl ester (40 m
g) was obtained. Melting point 169-174 [deg.] C. [Α] _D- 28.
3 ° (c0.28, CHCl ₃ ). FABMS (m /
z): 1482 (M + H) ⁺ . IR (KBr) ν: 34
00 (NH), 2100 (azide), 1740 (e
Ster), 1653, 1524 (amide). Elemental analysis: Found. C57.54; H7.55; N1
3.01. Calcd. C57.55; H7.62; N
13.23.

[0036]

Example 1 AZT (50 mg) was dissolved in THF (1 ml), p-nitrophenyl oxalyl chloride (108 mg) and triethylamine (48 mg) were added under ice cooling, and the mixture was stirred under an argon atmosphere for 18 hours. In this reaction liquid, D
L-prolyl-L-phenylalanine benzyl ester (100 mg) and 4-dimethylaminopyridine [hereinafter abbreviated as DMAP] dissolved in MF (4 ml) (12 m
g) was added and the mixture was stirred under an argon atmosphere for 24 hours. After concentration under reduced pressure, methylene chloride (100 ml) was added, and sodium bicarbonate water was added for 10
% Citric acid water, water, and brine, washed successively, dried over anhydrous magnesium sulfate, and concentrated. This was purified by silica gel chromatography (CHCl ₃ / MeOH = 10/1) and N-carbo- (3′-azido-3′-deoxythymidyloxycarbonyl) -L-prolyl-L-phenylalanine benzyl ester [1a] ( 40 mg) was obtained. Melting point 63-65 ° C. [Α] _D −24.6 ° (c
0.99, CHCl ₃ , 22 ° C). FABMS (m /
z): 674 (M + H) ⁺ . IR (KBr) ν: 342
0 (NH), 2100 (azide), 1744 (es)
ter), 1680, 1654 (amide). ¹ H-
NMR (CDCl ₃ ) δ = 9.27, 9.12 (1H,
eachbr s, NH), 7.73, 7.51 (1
H, each, H-6), 7.39 to 7.01 (1
0H, m, Ar), 6.40-6.28 (1H, m, H)
-1 '), 5.20 to 5.08 (2H, m, COOC H
₂ Ph), 4.94 to 4.85 (1H, m, Pro; N
CHCO), 4.56-4.21 (1H, m, Phe;
NHC H CO, H-3 ' , 5'), 4.15~4.11
(1H, m, H-4 ′), 3.84 to 3.60 (2H,
_{m, Pro; C H 2 NCH} ), 3.17~3.05 (2
H, m, Phe; CHCH ₂ Phe), 2.57-2.
31 (2H, m, H-2 ′), 2.29 to 1.76 (4
_{H, m, Pro; C H} 2 C H 2 CHCO), 1.92,
1.89 (3H, each br s, C H ₃ ).

[0037]

Example 2 AZT (50 mg) was dissolved in THF (1 ml), p-nitrophenyl oxalyl chloride (108 mg) and triethylamine (48 mg) were added under ice cooling, and the mixture was stirred under an argon atmosphere for 18 hours. In this reaction liquid, D
L-prolyl-D-phenylalanine benzyl ester (100 mg) and DMA dissolved in MF (4 ml)
P (12 mg) was added, and the mixture was stirred under an argon atmosphere for 24 hours. After concentration under reduced pressure, methylene chloride (100 ml) was added, the mixture was washed successively with aqueous sodium hydrogen carbonate, 10% aqueous citric acid, water and brine, dried over anhydrous magnesium sulfate and concentrated. This was subjected to silica gel chromatography (CHCl ₃ / MeOH = 10 /
Purified by 1), N-carbo- (3'-azido-3 '
-Deoxythymidyloxycarbonyl) -L-prolyl-D-phenylalanine benzyl ester [1b]
(40 mg) was obtained. Melting point 66-68 ° C. [Α] _D -3
8.2 ° (c0.99, CHCl ₃ , 22 ° C). FAB
MS (m / z): 674 (M + H) ^<+> . IR (KBr)
ν: 3460 (NH), 2100 (azide), 17
43 (ester), 1683, 1655 (amid
e). ¹ H-NMR (CDCl ₃ ) δ: 9.14, 8.
99 (1H, each brs, NH), 7.67,
7.49 (1H, each, H-6), 7.38-
6.97 (10H, m, Ar), 6.38-6.28
(1H, m, H-1 ′), 5.21 to 5.08 (2H,
m, COOC H ₂ Ph), 4.92 to 4.80 (1H,
m, Pro; NCHCO), 4.58 to 4.21 (1
H, m, Phe; NHC H CO, H-3 ', 5'),
4.13-4.10 (1H, m, H-4 '), 3.82
_{~3.56 (2H, m, Pro;} C H 2 NCH), 3.
18~3.02 (2H, m, Phe; CHC H 2 Ph
e), 2.55 to 2.27 (2H, m, H-2 '),
_{2.22~1.79 (4H, m, Pro;} C H 2 C H 2
CHCO), 1.92, 1.88 (3H, eachch b
r s, C H ₃ ).

[0038]

Example 3 ddU (50 mg) was dissolved in THF (1 ml), p-nitrophenyl oxalyl chloride (108 mg) and triethylamine (48 mg) were added under ice cooling, and the mixture was stirred under an argon atmosphere for 18 hours. In this reaction liquid, D
L-Prolyl-D-phenylalanine benzyl ester (127 mg) and DMA dissolved in MF (4 ml)
P (14 mg) was added, and the mixture was stirred under an argon atmosphere for 24 hours. After concentration under reduced pressure, methylene chloride (100 ml) was added, and the mixture was washed successively with aqueous sodium hydrogen carbonate, 10% aqueous citric acid, water and brine, dried over anhydrous magnesium sulfate and concentrated. This was subjected to silica gel chromatography (CHCl ₃ / MeOH = 10 /
1-purified, N-carbo- (2 ', 3'-dideoxyuridyloxycarbonyl) -L-prolyl-D-
Phenylalanine benzyl ester [1c] (36m
g) was obtained. Melting point 71-74 ° C. [Α] _D −42.8
° (c0.31, CHCl ₃ ). FABMS (m /
z): 619 (M + H) ⁺ . IR (KBr) ν: 348
6 (NH), 1743 (ester), 1666 (am)
ide). ¹ H-NMR (CDCl ₃ ) δ: 8.88,
8.82 (1H, each brs, NH), 7.9
8,7.87 (1H, each d, J = 4.6Hz,
H-6), 7.36 to 6.98 (10H, m, Ar),
6.18 to 6.11 (1H, m, H-1 '), 5.86
~ 5.76 (1H, eachch d, J = 4.6Hz, H
-5), 5.20 to 5.07 (2H, m, COOC H ₂
Ph), 4.92 to 4.72 (1H, m, Pro; NC
HCO), 4.56 to 4.19 (5H, m, Phe; N
CHCO, H-4 ′, 5 ′), 3.79 to 3.57 (2
_{H, m, Pro; C H} 2 NCH), 3.19~3.02
_{(2H, m, Phe; CHC} H 2 Phe), 2.46~
1.80 (8H, m, Pro; C H ₂ C H ₂ CHCO,
H-2 ', 3').

[0039]

Example 4 ddU (50 mg) in THF (1 ml),
Pyridine (23 mg) and DMAP (5 mg) were dissolved, and oxalyl chloride (0.
15 ml) and the mixture was stirred for 4 hours. The reaction solution was concentrated under reduced pressure, then L-prolyl-D-phenylalanine benzyl ester (127 mg) and pyridine (23 mg).
THF solution (1 ml) was added thereto, and the mixture was stirred under an argon atmosphere and ice cooling for 1 hour. After concentration under reduced pressure, purification by silica gel chromatography (CHCl ₃ / MeOH = 10/1), N-carbo- (5-chloro-2 ′, 3′-dideoxyuridyloxycarbonyl) -L-prolyl-D-phenylalanine benzyl was performed. Ester [1d] (18m
g) was obtained. Melting point 55-58 ° C. [Α] _D −42.3 °
(C0.20, CHCl _3). FABMS (m / z):
619 (M + H) ⁺ . IR (KBr) ν: 3456 (N
H), 1738 (ester), 1689 (amid)
e). ¹ H-NMR (CDCl ₃ ) δ: 8.56, 8.
48 (1H, each br s, NH), 8.09,
7.96 (1H, each, H-6), 7.37-
7.05 (10H, m, Ar), 6.11 to 6.06
(1H, m, H-1 ′), 5.19 to 5.09 (2H,
m, COOC H ₂ Ph), 4.96 to 4.81 (1H,
m, Pro; NCHCO), 4.57 to 4.20 (5
H, m, Phe; NCHCO, H-4 ', 5'), 3.
_{83~3.61 (2H, m, Pro;} C H 2 NCH),
3.16~3.03 (2H, m, Phe; CHC H 2 P
he), 2.44~1.79 (8H, m , Pro; C H
_{2 C H 2 CHCO, H-} 2 ', 3').

[0040]

Example 5 ddC (50 mg) was dissolved in DMF (1 ml), p-nitrophenyl oxalyl chloride (108 mg) and triethylamine (48 mg) were added under ice cooling, and the mixture was stirred under an argon atmosphere for 18 hours. In this reaction liquid, D
L-Prolyl-D-phenylalanine benzyl ester (127 mg) and DMA dissolved in MF (4 ml)
P (14 mg) was added, and the mixture was stirred under an argon atmosphere for 24 hours. After concentration under reduced pressure, methylene chloride (100 ml) was added, and the mixture was washed successively with aqueous sodium hydrogen carbonate, 10% aqueous citric acid, water and brine, dried over anhydrous magnesium sulfate and concentrated. This is subjected to silica gel chromatography (CHCl ₃ / MeOH = 6/1).
Purified by N-carbo- (2 ′, 3′-dideoxycytidyloxycarbonyl) -L-prolyl-D-phenylalanine benzyl ester [1e] (18 mg).
Got Melting point 72-76 ° C. [Α] _D −9.3 ° (c
0.20, CHCl _3). FABMS (m / z): 61
8 (M + H) ⁺ . IR (KBr) ν: 3452 (N
H), 1738 (ester), 1653 (amid
e). ¹ H-NMR (CDCl ₃ ) δ: 8.01 (1
H, br t, J = 7.6 Hz, H-6), 7.36-
6.98 (10H, m, Ar), 6.61 (2H, br
d, J = 7.6 Hz, NH ₂ ), 6.18 to 6.11
(1H, m, H-1 ′), 5.88 (1H, br t,
J = 7.6 Hz, H-5), 5.18 to 5.04 (2
H, m, COOC H ₂ Ph), 4.89 to 4.75 (1
H, m, Pro; NCHCO), 4.57 to 4.28.
(5H, m, Phe; NCHCO, H-3 ′, 4 ′,
5 '), 3.77~3.56 (2H, m, Pro; C H
₂ NCH), 3.18 to 3.04 (2H, m, Phe;
CHC H ₂ Phe), 2.44 to 2.06 (4H, m,
H-2 ', 3'), 2.01 to 1.58 (4H, m, P
_{ro; C H 2 C H 2} CHCO).

[0041]

Example 6 2 ', 3'-dideoxyinosine [hereinafter d
abbreviated as dI] (50 mg) dissolved in DMF (1 ml),
Under ice cooling, p-nitrophenyl oxalyl chloride (97m
g) and triethylamine (43 mg) were added, and the mixture was stirred under an argon atmosphere for 18 hours. DMF (4 m
l-prolyl-D-phenylalanine benzyl ester (113 mg) and DMAP (13 m)
g) was added and the mixture was stirred under an argon atmosphere for 24 hours. After concentration under reduced pressure, methylene chloride (100 ml) was added, and sodium bicarbonate water was added for 10 times.
% Citric acid water, water, and brine, washed successively, dried over anhydrous magnesium sulfate, and concentrated. This was purified by silica gel chromatography (CHCl ₃ / MeOH = 8/1) and N-carbo- (2 ′, 3′-dideoxyinosyloxycarbonyl) -L-prolyl-D-phenylalanine benzyl ester [1f] (28 mg ) Got. Melting point 75-77 ° C. [Α] _D −35.2 ° (c0.20,
CHCl ₃ ). FABMS (m / z): 643 (M +
H) ⁺ . IR (KBr) ν: 3428 (NH), 173
9 (ester), 1685 (amide). ¹ H-N
_{MR (CDCl 3) δ: 8.19} (1H, s, H-
2), 8.07 (1H, s, H-8), 7.35-7.
02 (10H, m, Ar), 6.82 (2H, brs,
_{NH 2), 6.28~6.25 (1H,} m, H-
1 ′), 5.18 to 5.06 (2H, m, COOC H ₂
Ph), 4.89-4.83 (1H, m, Pro; NC
HCO), 4.68-4.28 (3H, m, Phe; N
CHCO, H-5 '), 4.14-4.08 (1H,
m, H-4 ′), 3.72 to 3.51 (2H, m, Pr
o; C H ₂ NCH), 3.18 to 3.04 (2H, m,
Phe; CHC H ₂ Phe), 2.64 to 1.60 (8
_{H, m, Pro; C H} 2 C H 2 CHCO, H-2 ',
3 ').

[0042]

Example 7 ddA (50 mg) was dissolved in DMF (1 ml), p-nitrophenyl oxalyl chloride (97 mg) and triethylamine (43 mg) were added under ice cooling, and the mixture was stirred under an argon atmosphere for 18 hours. DM was added to this reaction solution.
L-prolyl-D-phenylalanine benzyl ester (113 mg) and DMAP dissolved in F (4 ml)
(13 mg) was added, and the mixture was stirred under an argon atmosphere for 24 hours. After concentration under reduced pressure, methylene chloride (100 ml) was added, and the mixture was washed successively with aqueous sodium hydrogen carbonate, 10% aqueous citric acid, water and brine, dried over anhydrous magnesium sulfate and concentrated. This is chromatographed on silica gel (CHCl ₃ / MeOH = 8/1).
And N-carbo- (2 ′, 3′-dideoxyadenosyloxycarbonyl) -L-prolyl-D-phenylalanine benzyl ester [1 g] (32 m
g) was obtained. Melting point 72-75 ° C. [Α] _D −2.4 °
(C0.20, CHCl _3). FABMS (m / z):
642 (M + H) ⁺ . IR (KBr) ν: 3460 (N
H), 1736 (ester), 1653 (amid
e). ¹ H-NMR (CDCl ₃ ) δ: 8.82 (1
H, s, H-2), 7.99 (1H, s, H-8),
7.33-7.14 (10H, m, Ar), 6.51-
6.48 (1H, m, H-1 '), 5.15 to 5.01
_{(2H, m, COOC H 2} Ph), 4.74~4.67
(1H, m, Pro; NCHCO), 4.55 to 4.2
8 (3H, m, Phe; NCHCO, H-5 '), 4.
06-3.83 (1H, m, H-4 '), 3.66-
3.35 (2H, m, Pro; C H ₂ NCH), 3.2
7~2.95 (2H, m, Phe; CHC H 2 Ph),
_{2.63~1.59 (8H, m, Pro;} C H 2 C H 2
CHCO, H-2 ', 3').

[0043]

Example 8 N-carbo- (3'-azido-3'-deoxythymidyloxycarbonyl) -O-tert-butyl-L-seryl-L-phenylalanyl-L-leucyl-O-tert-butyl- L-threonyl-N (ε)-
tert-Butoxycarbonyl-L-lysyl-glycyl-L-prolyl-O-tert-butyl-L-serine t
The ert-butyl ester (20 mg) was dissolved in trifluoroacetic acid (1 ml), and the mixture was stirred under an argon atmosphere for 1 hour. After concentration under reduced pressure at room temperature to dryness, ether was added to precipitate crystals. This was collected by filtration, vacuum dried and N-carbo-
(3′-Azido-3′-deoxythymidyloxycarbonyl) -L-seryl-L-phenylalanyl-L-leucyl-L-threonyl-L-lysyl-glycyl-L-prolyl-L-serine [1h] ( 30 mg) was obtained. Melting point 127-132 [deg.] C. [Α] _D −38.5 ° (c0.2
8, CHCl ₃ / MeOH = 1/1). FABMS (m
/ Z): 1158 (M + H) ⁺ . IR (KBr) ν: 3
388 (NH, OH), 2100 (azide), 17
40 (ester), 1662, 1534 (amid
e). Elemental analysis: Found. C52.05; H6.5
0; N17.20. Calcd. C51.90; H6.
27; N16.95.

[0044]

[Test Example 1] Examination of anti-HIV action The anti-HIV activity of the compound obtained in Production Example was measured by using MT-4 cells in an infection test system (T. Nagamo, H. Hos).
hino, Jpn. J. Cancer Res. , 7
9 , 9 (1988)). That is, HIV was added to the test substance, and 1 × 10 ⁵ MT-4 cells were infected with this. After incubating at 37 ° C. under 5% carbon dioxide gas for 4 days, smear of MT-4 cells was made, and the establishment of HIV infection was determined by the indirect fluorescent antibody method. The results are shown in Table 1 as a control when the test substance was not added and as a comparison with the control.

[0045]

[Table 1]

The values in the table show the percentage of cells that became HIV antigen-positive and became virus antigen positive. From the results in Table 1,
A clear anti-HIV effect was observed for the compounds of the present invention.

Claims (5)

[Claims] 1. The following general formula [1]: (In the formula, R ¹ is a hydrogen atom, R ² is an alkyl group, a hydroxymethyl group, or — (CH) _{n in which} R ¹ and R ² are connected.
-In n is 3 or 4, R ³ is a benzyl group which may have a substituent, X is a lower alkoxy, a benzyloxy group, an amino acid or peptide derivative, Y is a hydrogen atom or an azido group, and B is a nucleic acid. A peptide derivative of a dideoxynucleoside represented by (representing a base). 2. The following general formula [2]: The peptide derivative of dideoxynucleoside according to claim 1, wherein X, Y and B in the formula have the above-mentioned meanings. 3. The following general formula [3]: The peptide derivative of dideoxynucleoside according to claim 1, wherein X, Y and B in the formula have the above-mentioned meanings. 4. The following general formula [4]: The peptide derivative of dideoxynucleoside according to claim 1, wherein Y and B in the formula have the above-mentioned meanings. 5. An anti-HIV agent containing the peptide derivative of dideoxynucleoside according to claim 1 as an active ingredient.