CN102649788B - Beta-L-2'-desoxy-thymin-nucleoside derivative, preparation method and purposes thereof - Google Patents

Abstract

The invention belongs to the technical field of chemical medicines, and particularly relates to a Beta-L-2'-desoxy-thymine nucleoside derivative, a preparation method and purposes thereof. The Beta-L-2'-desoxy-thymine nucleoside derivative has a structure as showed in a format I, wherein R1 and R2 are independent O, S or Se; when one of the R1 and the R2 is O, R3 is halogen, -OH, -NH2, C1-C8 alkyl, (CH2)nF, (CH2)n-1CHF or CF3; R4 is halogen, H, or CF3; and n is 1-8. The invention provides the derivative which has activity of resisting hepatitis virus, and offers a new choice for preparing medicines capable of treating the hepatitis.

Description

β-L-2'-deoxy-thymidine derivatives and their preparation methods and uses

technical field

The invention belongs to the technical field of chemistry and medicine, and in particular relates to β-L-2'-deoxy-thymidine derivatives and their preparation methods and applications.

Background technique

Chronic hepatitis B (CHB) is a globally widespread infectious disease, and hepatitis B virus (Hepatitis B virus, HBV) infection is a great challenge to global public health. According to the statistics of the World Health Organization (WHO), among the approximately 2 billion people who have been infected with hepatitis B virus (HBV) in the world, more than 400 million people suffer from chronic hepatitis B virus infection, and about 1 million patients die of HBV every year. Liver cirrhosis or liver cancer associated with hepatitis virus infection (Kim KH, Kim ND, Seong BL. Molecules, 2010, 15(9):5878-5908). What's more serious is that 75% of the global hepatitis B virus infections are distributed in Asia. There are about 120 million hepatitis B virus-infected persons in my country, and the new cases of HBV each year exceed 3 million, and the annual treatment expenditure is as high as 30 billion to 50 billion yuan (Wang GF, Shi LP, Zuo JP. Virologica Sinica, 2008, 23 (2): 137-145). Therefore, research on anti-HBV drugs with independent intellectual property rights is an urgent social problem.

CHB often causes liver dysfunction, cirrhosis and hepatocellular carcinoma (Kremsdorf D, Soussan P, Paterlini-Brechot P, et al. Oncogene, 2006, 25: 3823-3833; Brechot C. Gastroenterology, 2004, 127: S56 -S61), the goal of antiviral therapy is to suppress or eliminate HBV to the greatest extent, improve the quality of life of patients and prolong their survival time. The current antiviral therapy is mainly to resist the virus and protect the body by intervening in the virus replication process or targeting different links of the host's anti-virus. The treatment of CHB includes drug therapy and non-drug therapy, among which drug therapy is dominant. Drug therapy includes interferon, thymosin, nucleoside drugs, non-nucleoside analogues, polypeptide drugs, therapeutic vaccines and Chinese herbal medicines and active ingredients. Among them, nucleoside drugs, as the first choice for the clinical treatment of viral diseases such as AIDS, hepatitis, and herpes, not only inhibit the enzymes of viral replication, but also participate in the competition and penetrate into the DNA of viral replication as substrate analogs, blocking DNA chain elongation, thereby inhibiting viral replication.

Telbivudine, as the fourth nucleoside analogue approved for marketing for the treatment of chronic hepatitis B virus, has the characteristics of good specificity and strong effect, and provides a new option for clinical practice. However, telbivudine has the following disadvantages: potential nephrotoxicity, drug resistance of the virus, the body's dependence on the drug, easy rebound after drug withdrawal, treatment must be started when the liver is onset, and some patients have rhabdomyolysis tendency etc. According to the bioelectronic isosteric principle, a fluorine atom is introduced into the 2' position of the sugar ring of telbivudine to obtain clavudine, which has better curative effect than telbivudine and lower toxicity than telbivudine, and can last after drug withdrawal. Inhibit HBV DNA for more than half a year (Jules L. Dienstag. Hepatology, 2009, 49(5): S112-S121). It shows that the drug designed by bioisosteric principle is an effective means to reduce drug toxicity and improve curative effect.

Contents of the invention

The first technical problem to be solved by the present invention is to provide derivatives of β-L-2'-deoxy-thymidine, the structure of which is shown in formula I:

Wherein, R ₁ and R ₂ are independently O, S or Se, and R ₁ and R ₂ are not O at the same time;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; n=1-8.

Preferably, R ₁ and R ₂ are independently O, S or Se, and R ₁ and R ₂ are not O at the same time;

R ₃ is F, Cl, Br, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; n=1-8.

Preferably, R ₁ and R ₂ are independently O, S or Se, and R ₁ and R ₂ are not O at the same time;

R ₃ is C ₁ ~ C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; n=1-8.

Preferably, R ₁ and R ₂ are independently O, S or Se, and R ₁ and R ₂ are not O at the same time;

R ₃ is C ₁ -C ₈ alkyl;

R ₄ is F, Cl, Br, H or CF ₃ .

Preferably, R ₁ and R ₂ are independently O, S or Se, and R ₁ and R ₂ are not O at the same time;

R ₃ is C ₁ -C ₈ alkyl;

_R4 is H.

Most preferably, R ₁ and R ₂ are independently O, S or Se, and R ₁ and R ₂ are not O at the same time;

_R3 is methyl; _R4 is H.

Further preferably, the derivative of β-L-2'-deoxy-thymidine has a structure as shown in formula II:

Wherein, R ₂ is independently S or Se;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; n=1-8.

Preferably, R ₂ is independently S or Se;

R ₃ is F, Cl, Br, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; n=1-8.

Preferably, R ₂ is independently S or Se; R ₃ is C ₁ -C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; n=1-8.

Preferably, R ₂ is independently S or Se; R ₃ is C ₁ -C ₈ alkyl; R ₄ is F, Cl, Br, H or CF ₃ .

Preferably, R ₂ is independently S or Se; R ₃ is C ₁ -C ₈ alkyl; R ₄ is H.

Preferably, R ₂ is independently S or Se; R ₃ is methyl; R ₄ is H.

Further preferably, the derivative of β-L-2'-deoxy-thymidine has a structure as shown in formula III:

Wherein, R ₂ is independently O, S or Se;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; n=1-8.

Preferably, R ₂ is independently O, S or Se;

R ₃ is F, Cl, Br, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; n=1-8.

Preferably, R ₂ is independently O, S or Se; R ₃ is C ₁ -C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; n=1-8.

Preferably, R ₂ is independently O, S or Se; R ₃ is C ₁ -C ₈ alkyl; R ₄ is F, Cl, Br, H or CF ₃ .

Preferably, R ₂ is independently O, S or Se; R ₃ is C ₁ -C ₈ alkyl; R ₄ is H.

Preferably, R ₂ is independently O, S or Se; R ₃ is methyl; R ₄ is H.

Further preferably, the derivative of β-L-2'-deoxy-thymidine has a structure as shown in formula IV:

Wherein, R ₂ is independently O, S or Se;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; n=1-8.

Preferably, R ₂ is independently O, S or Se;

R ₃ is F, Cl, Br, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; n=1-8.

Preferably, R ₂ is independently O, S or Se; R ₃ is C ₁ -C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; n=1-8.

Preferably, R ₂ is independently O, S or Se; R ₃ is C ₁ -C ₈ alkyl; R ₄ is F, Cl, Br, H or CF ₃ .

Preferably, R ₂ is independently O, S or Se; R ₃ is C ₁ -C ₈ alkyl; R ₄ is H.

Preferably, R ₂ is independently O, S or Se; R ₃ is methyl; R ₄ is H.

The second technical problem to be solved by the present invention is to provide intermediates used in the preparation of derivatives of β-L-2'-deoxy-thymidine shown in formula I, the structure of which is shown in formula V:

Wherein, R ₁ and R ₂ are independently O, S or Se;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; R ₅ and R ₆ are independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I, TBDMS, DMTr or Bz, and _R5 and _R6 are not H at the same time;

n=1-8.

Preferably, R ₁ and R ₂ are independently O, S or Se;

R ₃ is F, Cl, Br, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₅ and R ₆ are independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time; n=1-8.

Preferably, R ₁ and R ₂ are independently O, S or Se;

R ₃ is C ₁ ~ C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₅ and R ₆ are independently H, C1-C8 alkanoyl, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time;

n=1-8.

Preferably, R ₁ and R ₂ are independently O, S or Se;

R ₃ is C ₁ -C ₈ alkyl;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₅ and R ₆ are independently H, C1-C8 alkanoyl, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time.

Preferably, R ₁ and R ₂ are independently O, S or Se, and R ₁ and R ₂ are not O at the same time;

R ₃ is C ₁ -C ₈ alkyl;

R ₄ is H; R ₅ and R ₆ are independently H, C1-C8 alkanoyl, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time.

Most preferably, R ₁ and R ₂ are independently O, S or Se, and R ₁ and R ₂ are not O at the same time;

R ₃ is methyl; R ₄ is H; R ₅ and R ₆ are independently H, acetyl, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time.

Or the intermediate used during the preparation of derivatives of β-L-2'-deoxy-thymidine, the structure is as shown in formula VI:

Wherein, R ₁ is O, S or Se, R ₂ is Se;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; R ₅ and R ₆ are independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I, TBDMS, DMTr or Bz, and _R5 and _R6 are not H at the same time;

n=1-8.

Preferably, R ₁ is O, S or Se, R ₂ is Se;

R ₃ is F, Cl, Br, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₅ and R ₆ are independently C1-C8 alkanoyl, I, TBDMS, DMTr or Bz; n=1-8.

Preferably, R ₁ is O, S or Se, R ₂ is Se;

R ₃ is C ₁ ~ C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₅ and R ₆ are independently C1-C8 alkanoyl, I, TBDMS, DMTr or Bz;

n=1-8.

Preferably, R ₁ is O, S or Se, R ₂ is Se;

R ₃ is C ₁ -C ₈ alkyl;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₅ and R ₆ are independently C1-C8 alkanoyl, I, TBDMS, DMTr or Bz.

Preferably, R ₁ is O, S or Se, R ₂ is Se;

R ₃ is C ₁ -C ₈ alkyl;

R ₄ is H; R ₅ and R ₆ are independently C1-C8 alkanoyl, I, TBDMS, DMTr or Bz.

Preferably, R ₁ is O, S or Se; R ₂ is Se;

R ₃ is methyl; R ₄ is H; R ₅ and R ₆ are independently acetyl, I, TBDMS, DMTr or Bz.

Most preferably, R ₁ is O, S or Se; R ₂ is Se; R ₃ is methyl; R ₄ is H; R ₅ and R ₆ are Bz.

Or the intermediate used during the preparation of derivatives of β-L-2'-deoxy-thymidine, the structure is as shown in formula VII:

Wherein, R ₁ is O, S or Se;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; R ₅ and R ₆ are independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I, TBDMS, DMTr or Bz, and _R5 and _R6 are not H at the same time;

n=1-8.

Preferably, R ₁ is O, S or Se;

R ₃ is F, Cl, Br, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₅ and R ₆ are independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time; n=1-8.

Preferably, R ₁ is O, S or Se;

R ₃ is C ₁ ~ C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₅ and R ₆ are independently C1-C8 alkanoyl, I, TBDMS, DMTr or Bz;

n=1-8.

Preferably, R ₁ is O, S or Se;

R ₃ is C ₁ -C ₈ alkyl;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₅ and R ₆ are independently C1-C8 alkanoyl, I, TBDMS, DMTr or Bz.

Preferably, R ₁ is O, S or Se;

R ₃ is C ₁ -C ₈ alkyl;

R ₄ is H; R ₅ and R ₆ are independently C1-C8 alkanoyl, I, TBDMS, DMTr or Bz.

Preferably, R ₁ is O, S or Se;

R ₃ is methyl; R ₄ is H; R ₅ and R ₆ are independently acetyl, I, TBDMS, DMTr or Bz.

Most preferably, R ₁ is O, S or Se; R ₃ is methyl; R ₄ is H; R ₅ and R ₆ are Bz.

Or the intermediate used during the preparation of derivatives of β-L-2'-deoxy-thymidine, the structure is as shown in formula VIII:

Wherein, R ₁ is S, R ₂ is O, S or Se;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; R ₅ and R ₆ are independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time; n=1-8.

Preferably, R ₁ is S, R ₂ is O, S or Se;

R ₃ is F, Cl, Br, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₅ and R ₆ are independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time; n=1-8.

Preferably, R ₁ is S, R ₂ is O, S or Se;

R ₃ is C ₁ ~ C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₅ and R ₆ are independently H, C1-C8 alkanoyl, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time; n =1~8.

Preferably, R ₁ is S, R ₂ is O, S or Se; R ₃ is C ₁ -C ₈ alkyl;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₅ and R ₆ are independently H, C1-C8 alkanoyl, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time.

Preferably, R ₁ is S; R ₂ is O, S or Se; R ₃ is C ₁ -C ₈ alkyl;

R ₄ is H; R ₅ and R ₆ are independently H, acetyl, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time.

Preferably, R ₁ is S; R ₂ is O, S or Se; R ₃ is methyl; R ₄ is H; R ₅ is DMTr, and R ₆ is H.

Most preferably, R ₁ is S; R ₂ is O; R ₃ is methyl; R ₄ is H; R ₅ is DMTr, and R ₆ is H.

Or the intermediate used during the preparation of derivatives of β-L-2'-deoxy-thymidine, the structure is as shown in formula IX:

Wherein, R ₂ is independently O, S or Se;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; R ₆ is independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I, TBDMS, DMTr or Bz.

Preferably, R ₂ is independently O, S or Se;

R ₃ is F, Cl, Br, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₆ is independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I, TBDMS, DMTr or Bz.

Preferably, R ₂ is independently O, S or Se; R ₃ is C ₁ -C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₆ is independently C1-C8 alkanoyl, I, TBDMS, DMTr or Bz.

Preferably, R ₂ is O, S or Se; R ₃ is C ₁ -C ₈ alkyl;

R ₄ is F, Cl, Br, H or CF ₃ ; R ₆ is independently C1-C8 alkanoyl, I, TBDMS, DMTr or Bz.

Preferably, R ₂ is O, S or Se; R ₃ is C ₁ -C ₈ alkyl;

R ₄ is H; R ₆ is C1-C8 alkanoyl, TBDMS, DMTr or Bz.

Preferably, R ₂ is O; R ₃ is methyl; R ₄ is H; R ₆ is C ₁ -C ₈ alkanoyl.

Most preferably, _R2 is O; _R3 is methyl; _R4 is H; _R6 is acetyl.

The third technical problem to be solved by the present invention is to provide a preparation method for derivatives of β-L-2'-deoxy-thymidine shown in formula I, including the following:

one,

Wherein, R ₁ is O, S or Se;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; R ₅ and R ₆ are independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time; n=1-8.

Preferably, R ₅ and R ₆ are Bz.

Compound a obtains compound b through hydroxyl protection reaction, compound b is reacted under the catalysis of Lawesson's Reagent to obtain compound c, and compound c is obtained by removing the hydroxyl protecting group.

two,

Wherein, R ₁ is O, S or Se;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; R ₅ and R ₆ are independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I, TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time; n=1-8.

Preferably, R ₅ and R ₆ are Bz.

Compound b is reacted under the catalysis of POCl ₃ , 1,2,4-triazole, and Et ₃ N to obtain compound d, and compound d is reacted under the catalysis of NaBH ₄ and (NCCH ₂ CH ₂ Se) ₂ to obtain compound e, and compound e and NaOH The reaction of ethanol solution can be obtained.

three,

Wherein, R ₂ is O, S or Se;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; R ₅ is methyl, tert-butyl, 2-tetrahydropyranyl, C1~C8 alkanoyl, I, TBDMS, DMTr or Bz, R ₆ is H, methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, I, TBDMS, DMTr or Bz; n=1-8.

Preferably, R ₅ is DMTr, R ₆ is H.

Compound f is obtained by hydroxyl protection reaction to obtain compound g, compound g is reacted with iodomethane, DBU (1,8-diazabicyclo[5.4.0]undec-7-ene) to obtain compound h, after the reaction of selenium and _NaBH Compound i can be obtained by reacting with compound h, and compound i can be obtained by removing the hydroxyl protecting group.

Four,

Wherein, R ₂ is O, S or Se;

R ₃ is halogen, -OH, -NH ₂ , C ₁ ~C ₈ alkyl, (CH ₂ ) _n F, (CH ₂ ) _n-1 CHF ₂ or CF ₃ ;

R ₄ is halogen, H or CF ₃ ; R ₅ and R ₆ are independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I, TBDMS, DMTr or Bz; n =1~8.

Preferably, R ₅ is TBDMS, and R ₆ is C1-C8 alkanoyl.

Compound j obtains compound k through selective hydroxyl protection reaction, compound k reacts with methyl triphenoxyphosphorus iodide to obtain compound L, compound L reacts with silver acetate to obtain compound m, and compound m reacts with _H2S under alkaline conditions Compound n can be obtained through the reaction, and compound n can be obtained by deprotecting the hydroxyl group.

The fourth technical problem to be solved by the present invention is to provide the use of derivatives of β-L-2'-deoxy-thymidine shown in formula I in the preparation of drugs for treating hepatitis. The hepatitis is chronic hepatitis B or hepatitis C.

The pharmaceutical composition is prepared by adding common excipients to the active ingredient; the active ingredient is a derivative of β-L-2'-deoxy-thymidine shown in formula I, a pharmaceutically acceptable salt of the derivative, at least one of a hydrate of the derivative or a solvate of the derivative. It is preferably a derivative of β-L-2'-deoxy-thymidine shown in formula I.

The β-L-2'-deoxy-thymidine derivative of the present invention has anti-hepatitis virus activity, and provides a new option for preparing medicines for treating hepatitis.

Detailed ways

Example 14-Sulfur-2'-deoxy-L-thymidine

a. Synthesis of Compound 2

Take 9.68g of 2-deoxy-L-thymidine (1), add 200ml of anhydrous pyridine, stir to dissolve, add 9.6ml of benzoyl chloride, heat to 50°C for 10-20 hours, TLC detects that after the reaction is completed, Concentrate to about 30ml, add crushed ice while stirring, produce precipitate, filter, dry under pressure, and recrystallize with ethanol to obtain 14g of white solid.

¹ H NMR (400MHz, CDCl ₃ ): δ1.618(s, 3H), 2.308～2.382(m, 1H), 2.718(dd, 1H), 4.544(d, 1H), 4.667～4.828(m, 2H) , 5.662(d, 1H), 6.474(q, 1H), 7.472~8.082(m, 10H), 8.391(s, 1H).

b. Synthesis of Compound 3

Take 9g of compound 2, add 200ml of dioxane, stir to dissolve, add 15g of Lawesson's Reagent (Lawesson's Reagent), reflux reaction, TLC to monitor the reaction process, after the reaction is completed, concentrate to dryness, add water, extract with chloroform, and the water layer Concentrated to dryness, column chromatography gave 7.7g light yellow solid.

¹ H NMR (400MHz, DMSO): δ1.609(s, 3H), 2.504～2.682(m, 2H), 4.564～4.691(m, 3H), 5.661(d, 1H), 6.272(t, 1H), 7.508~7.723 (m, 7H), 7.991~8.048 (m, 4H), 12.808 (s, 1H).

c. Synthesis of Compound 4

Take 4.66g of compound 3, add 200ml of methanol, stir to dissolve, add 1.62g of sodium methoxide, and stir at room temperature. The reaction progress was monitored by TLC. After the reaction was completed, the pH was adjusted to neutral with glacial acetic acid, concentrated to dryness, and column chromatography gave 1.9 g of a yellow solid.

¹ H NMR (400MHz, DMSO): δ1.967(s, 3H), 2.127～2.155(q, 2H), 3.546～3.641(m, 2H), 3.796(q, 1H), 4.242(s, 1H), 5.101 (s, 1H), 5.273 (d, 1H), 6.107 (t, 1H), 7.896 (s, 1H), 12.704 (s, 1H).

Example 24-Selenium-2'-deoxy-L-thymidine

a. Synthesis of Compound 5

Add 33ml POCl ₃ dropwise to the anhydrous acetonitrile suspension (1000ml) containing 99g 1,2,4-triazole, protect with N ₂ , stir at 0°C for 10min; then add 200ml anhydrous Et ₃ N dropwise to the above mixture The solution was stirred at 0°C for 35 min; finally, 10.85 g of compound 2 in acetonitrile (500 ml) was added, and stirred at room temperature for 4 h. TLC detected that the raw material point disappeared, and about 200ml of saturated NaHCO ₃ was added, then extracted with ethyl acetate, washed with water, dried, and concentrated to give 5 (11g) as a white solid.

¹ H NMR (400MHz, CDCl ₃ ): 2.221(s, 3H), 2.309-2.382(m, 1H), 3.212(dd, 1H), 4.124(q, 1H), 4.675～4.725(m, 2H), 4.932 (dd, 1H), 5.682(d, 1H), 6.376(q, 1H), 7.401~7.661(m, 5H), 7.969-8.104(m, 5H), 8.215(s, 1H), 9.255(s, 1H) ).

b. Synthesis of Compound 6

1.5gNaBH ₄ was added to the round bottom flask containing 2.88ml (NCCH ₂ CH ₂ Se) ₂ and 120ml ethanol, in ice bath, under N ₂ protection was added to dissolve 6g of compound 5 in THF (50ml) solution, TLC detection reaction process . After the reaction was complete, about 150ml of water was added, the pH of the solution was adjusted to 7 with 10% acetic acid, concentrated to dryness, and column chromatography gave white solid 6 (5g).

¹ H NMR (400MHz, CDCl ₃ ): 1.784(s, 3H), 2.253-2.327(m, 1H), 2.980(t, 2H), 3.054-3.109(m, 1H), 3.334-3.421(m, 2H) , 4.657-4.691(m, 2H), 4.867(q, 1H), 5.651(d, 1H), 6.356(q, 1H), 7.263-7.497(m, 5H), 7.518-7.650(m, 2H), 7.966 -8.096 (m, 4H).

c. Synthesis of Compound 7

1g of sodium hydroxide was dissolved in 400ml of absolute ethanol, then 4.5g of compound 6 was added to the above NaOH ethanol solution, the reaction was stirred at room temperature, and the reaction progress was detected by TLC. After the reaction was completed, the pH of the solution was adjusted to 7 with 10% acetic acid, concentrated To dryness, add 50 ml of water to dissolve, extract several times with CH ₂ Cl ₂ , concentrate the aqueous phase to dryness, and perform silica gel column chromatography to obtain orange solid 7 (1.8 g).

¹ H NMR (400MHz, DMSO): δ2.087(s, 3H), 2.162(t, 2H), 3.560-3.650(m, 2H), 3.800(q, 1H), 4.239(s, 1H), 5.123( s, 1H), 5.280 (d, 1H), 6.038 (t, 1H), 8.048 (s, 1H), 13.301 (s, 1H).

Example 32-Sulfur-2'-deoxy-L-thymidine

a. Synthesis of Compound 8

Take 9.68g of compound 1, add 200ml of anhydrous DMF, stir to dissolve, add 8g of imidazole and 6g of tert-butyldimethylchlorosilane, nitrogen protection reaction, TLC detection After the reaction is completed, add 100ml of ethyl acetate, wash with water, sulfuric acid Magnesium was dried and concentrated to dryness to obtain 13.5 g of white solid.

¹ H NMR (400MHz, CDCl ₃ ): 1.118(d, 6H), 0.923(s, 9H), 1.921(s, 3H), 1.997(br, 1H), 2.080-2.148(m, 1H), 2.325-2.379 (m, 1H), 3.823-3.914(m, 2H), 4.029(q, 1H), 4.476(t, 1H), 6.362(q, 1H), 7.496(d, 1H), 8.222(s, 1H).

b. Synthesis of Compound 9

Take 11.9g of compound 8, add 100ml of dry pyridine, stir to dissolve, add 5ml of acetic anhydride, stir at room temperature for reaction, TLC detects that the reaction is complete, concentrate to dryness, and column chromatography to obtain 11.2g of white solid.

c. Synthesis of Compound 10

11.2 g of compound 9 was dissolved in 100 ml of THF, 8.8 g of tetrabutylammonium fluoride was added, the reaction was stirred at room temperature until the raw material disappeared, concentrated under reduced pressure to dryness, and 6.8 g of white solid was obtained by column chromatography.

¹ H NMR (400MHz, CDCl ₃ ): 1.935(s, 3H), 2.115(s, 3H), 2.355-2.436(m, 2H), 3.908-3.947(m, 2H), 4.102(d, 1H), 5.352 (t, 1H), 6.254(q, 1H), 7.501(s, 1H), 8.271(s, 1H).

d. Synthesis of Compound 11

Take 4.3g of compound 10 and 11g of methyl triphenoxyphosphorus iodide, add 50ml of DMF, stir at 25°C for 2 hours to complete the reaction, add 5ml of methanol, concentrate to dryness under reduced pressure, and obtain 5.5g of product by column chromatography.

¹ H NMR (400MHz, CDCl ₃ ): 1.952(s, 3H), 2.105(s, 3H), 2.098-2.407(m, 2H), 3.405-3.712(m, 2H), 4.011(m, 1H), 5.097 (m, 1H), 6.318(q, 1H), 7.576(q, 1H), 8.510(s, 1H).

e. Synthesis of Compound 12

Take 5g of compound 11 and 10g of silver acetate, add 200ml of dry acetonitrile, reflux reaction, TLC detects that the reaction is complete, cooling, suction filtration to remove silver acetate, concentration to dryness, column chromatography to obtain 2.8g of product.

f. Synthesis of Compound 13

Dissolve 2.2g of compound 12 in 200ml of DMF, add 25ml of triethylamine, cool to ~70°C, pass through H2S for reaction, after the completion of the reaction as detected by TLC, concentrate to dryness, and obtain 0.8g of product by column chromatography.

¹ H NMR (400MHz, CDCl ₃ ): 1.997(s, 3H), 2.142(s, 3H), 2.105-2.406(m, 2H), 3.401-3.720(m, 2H), 4.052(m, 1H), 5.101 (m, 1H), 6.314(q, 1H), 7.918(q, 1H), 9.451(s, 1H).

g. Synthesis of Compound 14

100mg of sodium hydroxide was dissolved in 50ml of absolute ethanol, then 600mg of compound 13 was added to the above NaOH ethanol solution, stirred and reacted at room temperature, TLC was used to detect the reaction process, after the reaction was completed, the pH of the solution was adjusted to 7 with 10% acetic acid, concentrated to After drying, add 50 ml of water to dissolve, extract several times with CH ₂ Cl ₂ , concentrate the aqueous phase to dryness, and perform silica gel column chromatography to obtain 14 (420 mg) as a white solid.

¹ H NMR (400MHz, DMSO): δ2.085(s, 3H), 2.204(t, 2H), 3.552-3.648(m, 2H), 3.815(q, 1H), 4.240(s, 1H), 5.136( s, 1H), 5.264 (d, 1H), 6.103 (t, 1H), 8.124 (s, 1H), 13.125 (s, 1H).

Example 42-Selenium-2'-deoxy-L-thymidine

a. Synthesis of Compound 15

Dissolve 5.2g of compound 14 and 100mg of DMAP in 100ml of dry pyridine, stir and dissolve at 0°C, add 9g of 4,4'-dimethoxytrityl chloride to the above solution, stir and react for 10 hours under nitrogen protection, and detect the reaction by TLC Complete, add 5ml of methanol, concentrate to dryness under reduced pressure, add 100ml of ethyl acetate, wash with water three times, 50ml each time, dry the organic phase with sodium sulfate, filter, concentrate to dryness, column chromatography gives 9.8g of colorless solid.

¹ H NMR (400MHz, CDCl ₃ ) δ: 1.454(s, 3H), 2.316(m, 1H), 2.685(ddd, 1H), 3.411(dd, 1H), 3.568(dd, 1H), 3.792(2s, 6H), 4.610(m, 1H), 6.851-6.853(m, 4H), 6.941(t, 1H), 7.428-7.536(m, 9H), 7.885(d, 1H), 8.653(m, 2H), 10.418 (br, 1H).

b. Synthesis of Compound 16

Dissolve 9g of compound 15 and 10ml of methyl iodide in 60ml of dry DMF, stir at 0°C under nitrogen protection, add 4ml of DBU to the above solution, stir at 0°C for 1 hour, then add 30ml of cold water to the reaction mixture, add 300ml of ethyl acetate ester. After washing with water, the organic layer was dried over sodium sulfate, filtered, concentrated to dryness, and 8.2 g of white solid was obtained by column chromatography.

¹ H NMR (400MHz, CDCl ₃ ) δ: 1.554(s, 3H), 2.342(m, 1H), 2.523(ddd, 1H), 2.581(s, 3H), 3.209(br, 1H), 3.415(dd, 1H), 3.547(dd, 1H), 3.785(2s, 6H), 4.136(m, 1H), 4.648(m, 1H), 6.241(dd, 1H), 6.752-6.896(m, 4H), 7.185-7.526 (m, 9H), 7.853 (s, 1H).

c. Synthesis of Compound 17

3.8g of selenium and 1.9g of NaBH4 were added to absolute dry ethanol (150ml) at 0°C, stirred for 30min, and 8g of compound 16 was dissolved in 100ml of absolute ethanol. Add the prepared sodium selenium hydride solution to the ethanol solution of compound 16, react at room temperature for 72 hours, add 300ml of ethyl acetate and 100ml of sodium chloride solution successively, wash the organic layer with saturated sodium chloride and water successively, dry over sodium sulfate, and filter. Concentrate to dryness and perform column chromatography to obtain 7.1 g of a yellow solid.

¹ H NMR (400 MHz, CDCl ₃ ) δ: 1.390 (s, 3H), 2.721-2.668 (m, 2H), 3.407 (dd, 1H), 3.570 (dd, 1H), 3.786 (2s, 6H), 4.141 ( m, 1H), 4.625(m, 1H), 6.842(m, 4H), 7.018(t, 1H), 7.134-7.396(m, 9H), 7.947(d, 1H), 10.634(br, 1H).

d. Synthesis of Compound 18

Take 7 g of compound 17, add 50 ml of trifluoroacetic acid/dichloromethane (2:8) solution, stir at room temperature until complete reaction, concentrate to dryness, and obtain 2.4 g of compound 18 by column chromatography.

¹ H NMR (400MHz, DMSO): δ2.182(s, 3H), 2.195(t, 2H), 3.543-3.658(m, 2H), 3.826(q, 1H), 4.237(s, 1H), 5.145( s, 1H), 5.270 (d, 1H), 6.109 (t, 1H), 8.140 (s, 1H), 13.146 (s, 1H).

Example 52, 4-dithio-2'-deoxy-L-thymidine

a. Synthesis of Compound 19

Take 7.8g of compound 14, add 200ml of anhydrous pyridine, stir to dissolve, add 7.5ml of benzoyl chloride, heat to 50°C and react for 10-20 hours, after the reaction is detected by TLC, concentrate to about 30ml, add crushed ice while stirring, A precipitate was formed, filtered, dried under pressure, and recrystallized from ethanol to obtain 12.5 g of compound 19.

b. Synthesis of Compound 20

Take 9.4g of compound 19, add 200ml of dioxane, stir to dissolve, add 14g of Lawson's reagent, reflux reaction, TLC monitors the reaction progress, after the reaction is completed, concentrate to dryness, add water, extract with chloroform, the water layer is concentrated to dryness, and the column Chromatography yielded 8.5 g of compound 20.

c. Synthesis of Compound 21

Take 7g of compound 20, add 200ml of methanol, stir to dissolve, add 1.8g of sodium methoxide, and stir at room temperature. The reaction progress was monitored by TLC. After the reaction was completed, the pH was adjusted to neutral with glacial acetic acid, concentrated to dryness, and column chromatographed to obtain 5.4 g of compound 21.

Example 62-Selenium-4-thio-2'-deoxy-L-thymidine

a. Synthesis of Compound 22

Take 9.2 g of compound 18, add 200 ml of anhydrous pyridine, stir to dissolve, add 7.2 ml of benzoyl chloride, heat to 50 ° C for 10 to 20 hours, TLC detects that the reaction is completed, concentrate to about 30 ml, add crushed ice while stirring, Precipitation occurred, filtered, dried under pressure, and recrystallized from ethanol to obtain 13.8 g of compound 22.

b. Synthesis of Compound 23

Take 10.3g of compound 22, add 200ml of dioxane, stir to dissolve, add 12g of Lawson's reagent, reflux reaction, TLC to monitor the reaction process, after the reaction is completed, concentrate to dryness, add water, extract with chloroform, the water layer is concentrated to dryness, and the column Chromatography yielded 8.2 g of compound 23.

c. Synthesis of Compound 24

Take 8g of compound 23, add 200ml of methanol, stir to dissolve, add 2g of sodium methoxide, and stir at room temperature. The reaction progress was monitored by TLC. After the reaction was completed, the pH was adjusted to neutral with glacial acetic acid, concentrated to dryness, and column chromatographed to obtain 3.6 g of compound 24.

Example 72, 4-diselenium-2'-deoxy-L-thymidine

a. Synthesis of Compound 25

Add 35ml POCl ₃ dropwise to the anhydrous acetonitrile suspension (1000ml) containing 100g 1,2,4-triazole, protect with N ₂ , stir at 0°C for 10min; then add 200ml anhydrous Et ₃ N dropwise to the above mixture The solution was stirred at 0°C for 35 min; finally, 11 g of compound 22 was added in acetonitrile solution (500 ml), and the reaction was stirred at room temperature. TLC detected that the raw material point disappeared, and about 200 ml of saturated NaHCO ₃ was added, then extracted with ethyl acetate, washed with water, After drying and concentration, 10.6 g of compound 25 was obtained.

b. Synthesis of Compound 26

_3gNaBH4 was added to a round-bottomed flask containing 6ml ( _NCCH2CH2Se ) ₂ and _200ml ethanol, ice bathed, and a solution of 10g compound 25 in THF (100ml) was added under the protection of _N2 , and the reaction progress was detected by TLC. After the reaction was complete, about 200ml of water was added, the pH of the solution was adjusted to 7 with 10% acetic acid, concentrated to dryness, and 9.5g of compound 26 was obtained by column chromatography.

c. Synthesis of Compound 27

2g of sodium hydroxide was dissolved in 800ml of absolute ethanol, then 9.2g of compound 26 was added to the above NaOH ethanol solution, the reaction was stirred at room temperature, and the reaction progress was detected by TLC. After the reaction was completed, the pH of the solution was adjusted to 7 with 10% acetic acid, concentrated To dryness, add 100ml of water to dissolve, extract several times with CH ₂ Cl ₂ , concentrate the aqueous phase to dryness, and perform silica gel column chromatography to obtain 3.7 g of compound 27.

Example 82-Sulfur-4-selenium-2'-deoxy-L-thymidine

a. Synthesis of Compound 28

Add 35ml POCl ₃ dropwise to the anhydrous acetonitrile suspension (1000ml) containing 100g 1,2,4-triazole, protect with N ₂ , stir at 0°C for 10min; then add 200ml anhydrous Et ₃ N dropwise to the above mixture In the solution, stir at 0°C for 35 min; finally add 10 g of compound 19 in acetonitrile solution (500 ml), stir the reaction at room temperature, TLC detects that the raw material point disappears, add about 200 ml of saturated NaHCO ₃ , then extract with ethyl acetate, wash with water, After drying and concentrating, 8.5 g of compound 28 was obtained.

b. Synthesis of Compound 29

3gNaBH ₄ was added to a round bottom flask containing 6ml (NCCH ₂ CH ₂ Se) ₂ and 200ml ethanol, in an ice bath, and under N ₂ protection, 9.5g of compound 28 was dissolved in THF (100ml) solution, and the reaction progress was detected by TLC. After the reaction was complete, about 200ml of water was added, the pH of the solution was adjusted to 7 with 10% acetic acid, concentrated to dryness, and 8.2g of compound 29 was obtained by column chromatography.

c. Synthesis of compound 30

2g of sodium hydroxide was dissolved in 800ml of absolute ethanol, then 8g of compound 29 was added to the above NaOH ethanol solution, stirred and reacted at room temperature, TLC was used to detect the reaction process, after the reaction was completed, the pH of the solution was adjusted to 7 with 10% acetic acid, concentrated to After drying, add 100ml of water to dissolve, extract several times with CH ₂ Cl ₂ , concentrate the aqueous phase to dryness, and perform silica gel column chromatography to obtain 3.6 g of compound 30.

Test example 1 in vitro anti-HBV activity

a. Inhibitory effect on HBV DNA

Take the HepG2.2.15 cells in the logarithmic growth phase, adjust the cell concentration to 1×10 ⁴ /ml with DMM medium containing 10% fetal bovine serum, add 100 μL of single cell suspension to a 96-well culture plate, and store at 37°C, Cultured under the condition of 5% CO ₂ , and when the cells grew to confluence, 100 μL/well of compounds of different concentrations were added to the experimental group. The positive control group was added with different concentrations of telbivudine and continued to culture. The culture solution was taken out on the 3rd and 6th day, and the same volume of medicinal solution was added at the same time, and the culture solution was taken out on the 9th day and placed at ~20°C for use. At the same time, the same volume of culture medium was added for the determination of MTT, and whether the drug was toxic to the cells at each experimental concentration was examined.

b. Cytotoxicity test

Except for the difference in drug concentration, other operating steps are the same as part a, which is used to determine the half-toxic concentration (CC ₅₀ ) of the drug on cells. The results are shown in Table 1:

Table 1

The anti-HBV activity of the drug was evaluated by the Selectivity Index (SI), and the SI was calculated according to the following formula:

SI= _CC50 / _EC50

When SI>2, it indicates that the drug is effective and low toxicity; when 1≤SI≤2, it indicates that the drug is effective and toxic; when SI<1, it indicates that the drug has toxic effects. The larger the SI value, the stronger the inhibitory effect of the compound on HBV and the smaller the cytotoxicity.

As can be seen from the results in Table 1, the obtained 8 nucleoside analogs have a good inhibitory effect on the secretion of HBV DNA by HBV2.2.15 cells, and the selection index is all much greater than 2, and the selection index (260333) of one of the compounds 4 is far greater than that of the listed drug The selection index of telbivudine (91000) has a good application prospect.

Claims (5)

1. β-L-2'-deoxy-thymidine derivatives, characterized in that: the structure is as shown in formula II: _R2 is S; _R3 is methyl; _R4 is H. 2. The intermediate used during the preparation of derivatives of β-L-2'-deoxy-thymidine shown in formula II, the structure is as shown in formula V: Among them, R ₁ is O, R ₂ is S; R ₃ is methyl; R ₄ is H; R ₅ and R ₆ are independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkane Acyl, H, I, TBDMS, DMTr or Bz, and _R5 and _R6 are not H at the same time. 3. the preparation method of the derivative of β-L-2'-deoxy-thymidine shown in claim 1 formula II: Among them, R ₁ is O; R ₃ is methyl; R ₄ is H; R ₅ and R ₆ are independently methyl, tert-butyl, 2-tetrahydropyranyl, C1-C8 alkanoyl, H, I , TBDMS, DMTr or Bz, and R ₅ and R ₆ are not H at the same time; Compound a obtains compound b through hydroxyl protection reaction, compound b is reacted under the catalysis of Lawson's reagent to obtain compound c, and compound c is obtained by removing the hydroxyl protection group. 4. the purposes of the derivant of β-L-2'-deoxy-thymidine described in claim 1 in the preparation treatment hepatitis medicine. 5. The pharmaceutical composition is prepared by adding the commonly used adjuvant of the drug to the active ingredient; the active ingredient is the derivative of β-L-2'-deoxy-thymidine according to claim 1 or the drug of the derivative. Use at least one kind of salt.