CN1068035A - Use double-stranded RNA to eliminate the drug resistance of virus to nucleoside analog - Google Patents

Abstract

Early stage or late period of treatment of infection promptly when the gene mutation of virus taken place with recovery to otherwise when being the sensitivity of invalid antiviral agent, take wrong paired dsRNA, reduce with in the nucleoside analog antiviral agent therapeutic process drug resistance development speed of virus.Use wrong paired dsRNA,, but especially obviously reach the appearance of postponing and/or reducing appearance, especially HIV in the CD4 lymphocyte the retroviral of nucleoside analogue drug-resistant at the periphery blood mononuclear cell.

Description

Use double-stranded RNA to eliminate the drug resistance of virus to nucleoside analog

Nucleoside analog is usually as antiviral agent, especially as anti-reversibility virus (retroviruses) agent.Virus experience genovariation or sudden change cause the relative drug resistance of these antiviral agent.In case the generation drug resistance, virus is bred rapidly, so quickened potential seizure of disease.The early deployment double-stranded RNA (dsRNA) that infects can reduce the speed that viral drug resistance occurs.Even infect the later stage, gene mutation takes place, and dsRNA also can recover the sensitivity of virus to other invalid antiviral agent.This point can be combined by two kinds of patterns of life-time service clinically and find out in the unpredictable consequence that treatment causes.This clinical effectiveness is, host immune function recovers greatly, and detectable virus is very little.This result uses any one antiviral agent unexistent separately.

The long-term treatment of viral disease, particularly reversible viral disease relevant (list of references 1 and 2) occur with viral Drug resistance.Ideal situation is that when using nucleoside analog to carry out chemotherapy, nucleoside can be inserted among the viral gene information, and causes genetic flaw or incomplete, thereby reduces the viral growth cycle efficieny.The ability that defective or incomplete viral offspring forms infection reduces.But, by modifying the gene structure of itself, drug resistance can appear in virus relatively, thereby even there is nucleoside analog, also can produce infective viral offspring.Typical gene variation appears at pol gene (i.e. the virus component that instructs anti-viral nucleoside to insert in the growth circulation first step), and making virus can be subjected to antiviral to block form influences.But best research example is the interaction between retroviral, for example HIV(HIV (human immunodeficiency virus) at present) and 3 '-azido-3 '-deoxythymidine, the latter also is referred to as AZT or Zidovudine.Cause that AIT is had drug-fast mutation usually simultaneously to other medicines, for example didanosine (DDI) and zalcitabine (DDC) (being not limited to this two kinds of chemical compounds) drug resistance.

Had been found that before using the nucleoside analog treatment use the dsRNA of effective dose erroneous matching at reasonable time, the patient that can make HIV infect (the HIV positive) postpones or reduces, or not only postponed but also reduce the program that nucleoside analogue drug-resistant virus occurs.These programs make patient infect the later stage at HIV, when needing to use the nucleoside analog antiviral agent, to medicaments insensitive.

Also describe the dsRNA that uses the effective dose erroneous matching, but improved retrovirus infection patient's peripheral blood monokaryon hemocyte (PBMC) group, especially T ₄Or CD ₄Lymphocytic treatment procedure.

DsRNA contains the polyinosine of part uracil base or guanyl-and the complex of poly-cytidine, and for example the content of this class alkali base is from 1/4 to 1/29(poly I.poly(C _4-29X＞U or G)).

The general formula of dsRNA can be rI _nR(C _{11-14 '}U) _nOr rI _nR(C _{12 '}U) _nOther suitable example of dsRNA is discussed below.

The hydrogen bond (base stacking) that " the paired dsRNA of mistake " is used for describing between those marriage chains is complete relatively, and promptly hydrogen bond is interfered interrupted average in successive per 29 base pair residues during 1 base of less than." the paired dsRNA of mistake " should understand by this.

The present invention advocates the wrong paired dsRNA of use to be selected from poly(C _{N '}U) and poly(C _{N '}G) copolymerization nucleotide is the basis, and wherein n is 4 to 29 integer, and copolymerization nucleotide is by the poly-ribose cytidine key (rC in edge _n) rI _nRI _nModify and insert not timing base (uracil or guanine) and the wrong paired analog of the complex of the poly-ribose inosinic acid that generates and poly-ribose cytidylic acid.In addition, by modifying poly-ribose inosinic acid (rI _n) the ribose skeleton, for example 2 '-O-methylribose residue is included in wherein, from poly(I) poly(C) the dsRNA wrong paired dsRNA that derives.The paired complex of mistake can use the high polymer of RNA-stabilisation, and for example polylysine or cellulose carry out compound.Carter and Ts ' o be at United States Patent (USP) 4,130, described rI in 641 and 4,024,222 _nRC _{N '}Mistake pairing analog in preferred analog, general formula is rI _n(C _{11-14 '}U) _nOr rI _nLr(C _{29 '}G) _{N '}, list of references is classified in these inventions as.According to the present invention, wherein the dsRNA of Miao Shuing generally is suitable for.Preferred wrong paired dsRNA is rI _n(C _{11-14 '}U) _nOr AMPLIGEN(HEN Resarch, Inc, Rockn ille, MD USA), can buy lyophilized powder.

Other example of the wrong paired dsRNA that uses among the present invention comprises:

poly（I）·poly（C _4′，U）

poly（I）·poly（C _7′，U）

poly（I）·poly（C _13′，U）

poly（I）·poly（C ₂₂，U）

poly（I）·poly（C ₂₀，U）

Poly(I) poly(C ₂₉, U) and

poly（I）·poly C _P23G＞p

Be applicable to that implementing another kind of dsRNA of the present invention is the short chain dsRNA that determines structure, for example general formula is

5 '-block-(I) _n-blockade 3 '

3 '-block-(C) _n-blockade 5 '

Oligonucleotide.Wherein all greater than 5 to less than 100, I is single phosphorylation inosine separately for m and n, and C is single phosphorylation cytidine, and the blockade base complementation on the blockade base on chain and the relative chain.Or general formula is

5 '-block-((I) _xA) _j-blockade 3 '

5 '-block-((C) _yU) _k-blockade 3 '

Oligonucleotide.Wherein x and y be separately greater than 5 less than 25, and j and k equal 1 and less than 10 at least, and I and C are the same, and A is the nucleotide beyond the I, and U is base and the paired nucleotide of A.

In addition, the short chain oligonucleotide can have following structure:

5 ' (I) _n-hinge-(C) _m3 '

N wherein, m, the definition of I and C is the same.

These oligonucleotide can with relative chain on the not complementary chain of nucleotide on contain substituent group.These oligonucleotide are preferably stablized by the pairing of the inside of complementary non-homogeneous high polymer, and ideally, blockade base and hinge base or the two all contain the zone of complementary non-homogeneous high polymer.These oligonucleotide contain single last-of-chain ideally.These oligonucleotide are existing the description in PCT/US89/02172.

Patient need be according to clinical improvements situation defeated 200 to 700mg rIr(C of vein once in a week or once a day _{11-14 '}U).The consumption of dsRNA and administration number of times will make patient's systemic blood circulate in to be measured to the dsRNA level in the place away from transfusion part after the medication and reach 0.01 μ g to 1,000 μ g/ liter.

Illustrative nucleoside analog antiviral agent comprises that Ziodovudine(resembles AZT or zidovudine pyridine or the Retrovar that generally uses here), promptly 3 '-nitrine-3 '-the deoxidation thynlidine, be to be used for because of HIV (human immunodeficiency virus) (HIV, HTLV-I, the HTLV-II, HTLV-III, LAV, the similar information thing of ARV and various bacterial strains) the relevant complex situations with AIDS of AIDS that cause need the nucleoside analog antiviral agent of systemic treatment.Adult's dosage is administration 200mg per four hours of every day.The patient of a 70kg body weight, corresponding dosage is per four hours per kilogram of body weight 2.9mg.Every day per kilogram of body weight dosage with reaching 60mg.To show further that as the discussion of back during with the paired dsRNA coordination of mistake, the reversible viral dosage of the nucleoside analog of use is usually less than normal dose and custom consumption.

Fig. 1 is time (month number) table of describing 298 patient deaths or AIDS " systemic infection ", back AZT " late period " (circle) occurs with AZT " in early days " (grid) treatment before symptom occurs or symptom and treat and compare, adopt the patient's of no fatal HIV situation ratio to represent.

Fig. 2, but use dsRNA or it and AZT more in early days and use to the control of reversible viral growth with use retroviral ramp in the AZT case separately.In Fig. 2, HIV cultivates and is undertaken by list of references 3, and the PBMC of use is from example ^#1 to example ^#3 cultivated 4 days, after 7 days and 14 days, collected PBMC, be dissolved among the GUSCN, and by the use molecular hybrid technical measurement HIVRNA that describes in the method.Mononuclear cell in the PBMC=peripheral blood; GUSCN is a cyanic acid guanidine solution, is used for dissolved cell and releasing virus material.

Fig. 3, when relatively dsRNA and AZT are as the single therapy agent during with placebo and dsRNA and AZT therapeutic alliance, CD in the long-term maintenance HIV disease on indicated all numbers ₄Cell is pressed the relative effect of the middle qualitative changeization in the CD4T lymphocyte.

Fig. 4 is associated the natural law of AZT and dsRNA therapeutic alliance with the percent change of T4 level, show dsRNA Ampligen

T in the HIV disease ₄Expand to the stable period that has exceeded independent use AZT expection the stable period of cell.Pointed out each member's of drug combination middle dosage.

Fig. 5 is Ampligen

Natural law and T with the AZT coupling ₄The average percentage of cell changes (increase) and is associated, and shows Ampligen

T in increase and/or the stable HIV disease ₄Cellular level still exceeds this time cycle at AZT in the effective time cycle.

Fig. 6 be 12 the middle of the month placebo, use AZT, Ampligen separately

Single therapy and Ampligen

There is not fatal patient's HIV ratio diagram during with the AZT therapeutic alliance.

In Fig. 4 and Fig. 6, indicate near the bracket of patient's number relevant data point, in Fig. 3 and Fig. 5, indicate with N=.

In Fig. 2, according to the method for describing in the list of references 3, use is finished HIV from the peripheral blood mononuclear cell of example 1 and 3 and is cultivated altogether.Cultivated altogether 4 days, and after 7 days and 14 days, collected the peripheral blood mononuclear cell and also be dissolved in the cyanic acid guanidine solution, the latter is used for dissolved cell and releasing virus material.Measure HIV RNA with molecular hybrid, will describe in detail below.

Fig. 1 explanation is (for example weightless at " in early days " of the Veterans of the abundant announcement of hiv infected patient group, before symptoms such as night sweat occur) or " late period " (after comprising that promptly this class symptom by the relevant infection of fungus or bacterial HIV occurs) when using the AZT treatment, the development of fatal situation (developing into the AIDS or the death of systemic infection).

Can find out obviously that from Fig. 1 " in early days " uses not life-saving of AZT.The blood sample of taking from the patient of having developed fatal situation shows the AZT drug-resistant virus, and (this paper is with AZT

Expression) concentration increases, and the patient who did not still have fatal situation in about 40 months observation cycles generally has couple AZT to become responsive virus (AZTS).

Below, feature is more fully: although treat from AZT, the relative AZT sensitivity of the isolating on one's body typical HIV chorista of the patient that disease still develops, and these results are compared with comprising the viral chorista that obtains the hepatitis virus that the patient of relative good control is separated on one's body from disease.Clearly, AZT appears Virus can shorten the vital stage.But typical human body retroviral HTLV-1, HTLV-2, HTLV-3 and the virus of breeding by same mechanism comprise some hepatitis viruss.

The invention that improves this present an open question is described this inventor.This invention can be implemented by number of ways, and these approach comprise: (a) before virus is exposed to AZT or other analog, use the dsRNA of specific type, reduce AZT

Appearance (b) by the anti-dsRNA that adds in system, overcome AZT HIV(or other viral drug resistance analog) deadly character.DsRNA/ nucleoside system has shown antagonism AZT

The unexpected synergism of HIV and not have corresponding collaborative toxicity, this like this system be unique unexpected useful combination medicine really, use correctly can be saved life.

The inventor's study tour how to unite and use Ampligen

-AZT reduces the problem that HIV-virus is caused, in the research virus acellular from or accept Ampliger Treat separately or accept AZT and treat or accept Ampligen separately

The patient of-AZT therapeutic alliance, this virus is used to infect fresh human peripheral liquid cell, and this blood blood born of the same parents are exposed on Ampligen in advance

, AZT or Ampligen

Among AZT.Come from only accept the patient of AZT single therapy more than 1 year virus to insensitive (the 0.5uMAZT=4% suppression ratio of AZT; ED ₅₀＜5uM), but as original HIV virus to single Ampligen sensitivity (5ug/ml, Ampligen

-AZT coupling is than single Ampligen

Can produce bigger HIV suppression ratio (93%).Come from and accept single Ampligen

Treatment or Ampligen The patient's of-AZT therapeutic alliance virus has shown the relative drug resistance to AZT, to the Ampligen sensitivity

, and to Ampligen -AZT drug combination even bigger sensitivity is arranged.According to patient's different treatment plans, relatively use the result of many different virus chorista gained, can obviously see, before being exposed to AZT, just accepted Ampligen

The treatment patient virus for AZT(and other nucleoside analog) sensitivity be greater than the virus that comes from the patient who has at first accepted AZT.

A=AZT-Ampligen Interact by waiting number assay determination (list of references 4).Association index (CI) is defined as CI+(Ac/As)+(Be/Bs), wherein Ac and Bc are the drug level that uses in the therapeutic alliance, As and Bs are the drug level that produces the therapeutic alliance same effect when using separately.When CI＜1, medicine is worked in coordination with; When CI=1, medicine add and; When CI＞1, the mutual antagonism of medicine.ED ₅₀For suppressing the external dosage of 50% virus breeding.

The inventor shows typical embodiment of the present invention in Fig. 2, uses Ampligen in embodiments in early days

Or use simultaneously with AZT, in fact can produce substantial benefit, but resemble the bulk concentration of the so harmful virus of retroviral such as reduction.The generation curve of the rapid rising of the viral RNA of representing with open loop (ribonucleic acid) is an indication of not controlling viral growth, and two horizontal lines are represented to produce by therapeutic alliance virus and are well controlled.Open loop explanation AZT among Fig. 2

Virus develops rapidly, so just becomes the classical symptom of patient among Fig. 1, and these patients are when giving AZT separately, toward the development in more late period of disease.

Drug resistance Phenotype (table 1) whatsoever, some dsRNA, especially wrong paired dsRNA if with AZT(or other nucleoside analog) coupling, but then show coordinate repression to retroviral.H112-2 and 6910-6 are the HIV phenotype clones who shows typical sensitivity and drug-fast very characterization respectively.And example ^#1 to example ^#The 6th, (for example routine from being exposed to various systems ^#1,2,3 patients are identical with patient among Fig. 2, and for example 4,5,6 patients resemble and advise among Fig. 1, at first treat separately with AZT) especial patient in isolated virus.

When drug combination clinically, the result is unexpected once more, and not measurable according to the clinical observation of two kinds of medicines of independent use (single therapy).For example, use AZT(and other analog separately) make some immunocytes increase (list of references 5) in short-term, these immunocytes are called CD ₄Cell, but they the easiest be some retrovirals and some hepatitis viruss (particularly HHV6) attack.After increasing in short-term (seeing about 12 to 16 weeks), because HIV propagation and AZT occurs

HIV, immunocyte worsens (Fig. 3).Number in the bracket is called the main body number of research.Outside comparing, independent cooperation Ampligen All the time cause CD ₄Cell number is in horizontal line (Fig. 3, for example immuning cell number neither increases also and do not reduce, in other words, cell number stabilisation), Ampligen

Dosage about 400 to 500mg, weekly twice, intravenously administrable.

Patient accepts AZT and treats separately, oral 200mg(1200mg/ day of per four hours of every day).Ampligen

The independent patient of treatment group twice of the minimum dosage of vein (mean dose scope for 463-555mg) weekly weekly.(patient who accepts therapeutic alliance who shows among Fig. 4 also fails with 400mg typical A mpligen for twice weekly

Dosage, and the AZT of average companion's usefulness dosage range every day 300 to 540mg).

Placebo conforms to the evidence that disease process unfeelingly fails with the trend that AZT single therapy group (after 16 weeks) T4 lymphocyte intermediate value changes.Effectively treatment is not intervened, and placebo patient's lymphocytic intermediate value change count descends.After at first increasing, though accept the patient of AZT single therapy also experience they T4 cell intermediate value counting decline and AZT appears

HIV.This decline parallels since the decline that occurred in the 12nd week occurring among the patient with placebo treatment.

Treat separately and placebo group 12 week beginning T according to AZT ₄The linear regression line statistical method that the cell intermediate value change to constitute is slope relatively, shows they and intermediate value T ₄Unconverted 0 slope of cellular level has statistics difference (p＜0.01 and p＜0.01 is arranged respectively)." no change " typically represents the stable of amynologic disease, " negative slope " expression disease progression, and optimal consequence to be positive section lead, expression disease (immunity) is recovered, but this can not reach by the single therapy system of research.

The AZT single therapy of placebo is after 12 weeks, Ampligen

Eliminated the CD that expects separately ₄Descend.Ampligen

Continuous intermediate value CD in the treatment ₄The slope of regression line of level and the no CD of reflection ₄The horizontal line of loss cell does not all have significant difference in the whole time.

Slope is compared with horizontal line (no change)

The P value

Ampligen

Not statistically significant P〉0.2

AZT significant difference P＜0.01

Placebo significant difference P＜0.01

Though Ampligen Single therapy (Fig. 3) can eliminate the serious decline of the intermediate value T4 lymphocyte of in two groups of patients of placebo and AZT treatment, all seeing (after 12 weeks in short-term raise), but average T 4 levels do not raise.And at Ampligen

In the group, observed little (no difference of science of statistics) intermediate value T4 lymphopenia (Fig. 3) can be produced more persistent T4 stabilisation and reverse easily more than twice 200mg by dosage being increased to weekly in 1 year.At Ampligen

In the single therapy group since 12 weeks observed intermediate value T4 cells variation formation the regression line slope and show stable disease 0 between do not have significant difference.The Ampligen that Fig. 3 provides

Single therapy group, AZT single therapy group or placebo treatment group have about equally intermediate value and average absolute T4(also claim CD4) lymphocyte level.Then I they with accept Ampligen

Compare with new group of AZT therapeutic alliance.This new group abswolute level that in fact has the prognosis indication of lower (about 33%) intermediate value and this HIV infection that increases the weight of, thus show that they have bigger death risk or other " fatal event ".

Following table has provided Ampligen

With every mm in the AZT long term maintenance HIV disease ³The relative effect of cd4 cell.

As what in Fig. 3, see, finish 59 cells of the absolute T4 lymphocyte count minimizing of patient's intermediate value (4.9 cells of minimizing in about every month) that placebo is used in treatment in a year; The patient who accepts the AZT single therapy reduces 28 cells (2.3 cells of minimizing in about every month); Accept Ampligen

The patient of single therapy only reduces 15 cells (average every month reduce approximately 1.25 cells).But, accept Ampli-gen

With AZT therapeutic alliance group, this group has minimum pretreat intermediate value and average absolute T4 lymphocyte level (so being among the maximum risk), in fact having experienced the absolute T4 cellular level of intermediate value has increased by 13 cells, even after the therapeutic alliance 1 year, still like this.These results are summarised in the table 2, and are listed as more in detail among Fig. 4.

Fig. 4 has illustrated intermediate value T in a year of therapeutic alliance ₄The continuous variation of lymphocyte level, this therapeutic alliance should with the Ampligen among Fig. 3

Single therapy, AZT single therapy and placebo effect are compared.The patient that three month immediately measure of the baseline that is used for comparison before continuously absolute T according to the beginning therapeutic alliance ₄The meansigma methods of lymphocyte count is calculated.

In 90 day cycle among Fig. 3 of observed increase and original report the preliminary effect of observed AZT single therapy conform to.But, that is seen continued to increase the demonstration significant difference from 180 days to 540 days the therapeutic alliance, and this does not observe in the patient who accepts the AZT single therapy.(the nonparametric analysis has confirmed T in a year ₄Counting increases, and has statistical significance (p＞0.05).Why carrying out this analysis, is because relatively little sample number makes that possible T horizontal distribution is undesired).So, T ₄It is longer than 3 to 6 months increase in short-term of AZT one single treatment expectation that lymphocyte increases the time that continues, and this result must come from the existence of dsRNA.In addition, when using the AZT of relative low dosage and dsRNA, also see this effect.This observation and being consistent of previously under the situation that is low to moderate 200mg dosage, seeing.Intravenous injection twice weekly, continues 1 to 4 months, Ampligen

Single therapy can stablize untreated hiv infected patient (T4=60-300 cell/mm ³) the T4 cell of expectation reduce, if patient continued this treatment 5 to 8 months, can keep this effect (Fig. 3).

According to T in dsRNA and AZT therapeutic alliance 135 days to 630 days ₄The average percentage of lymphocyte count changes, and the regression line that obtains sees Fig. 5.The average of each patient's continuous T lymphocyte count in 91 to 180 days of therapeutic alliance is as the baseline of determining that percentage changes.This baseline is selected as separation, and in the back of this point, the AZT of estimation is to T ₄The influence of level generally disappears.

Use statistical method, the slope of the positive slope of the regression line and 95% credible limit is pointed out Ampligen

Successfully stoped the untreated patient (4-6 cell/moon) that expects and long-term AZT single therapy patient's (more than 6 cell/moons) long-term T with the AZT therapeutic alliance ₄Cell reduces.So, T has in fact been kept in therapeutic alliance indefinitely ₄Level, and the persistent period is observed more much longer than original AZT single therapy, and this point has just in time illustrated the present invention at the nucleic acid drug-resistant virus, the basic purposes in the particularly reversible viral therapy.

At last, Fig. 6 accepts placebo, AZT single therapy, Ampligen by statement as patient

Single therapy, perhaps Ampligen

During with the AZT therapeutic alliance, develop into the ratio of AIDA infection/lymphoma (" fatal event "), confirmed effectiveness of the present invention.

Point out that as Fig. 6 accepting has about 10% experience fatal event in 12 months observation cycle among the patient of placebo.Patient's number of beginning, 6th month and 12nd month of research indicated in bracket.In this period, accept to have among the patient of AZT about 4% experience fatal event.Should be pointed out that the tumor (list of references 6) that independent use AZT can expect formation especially, be avoided by therapeutic alliance.Use Ampligen

During with the AZT therapeutic alliance, at absolute T ₄Lymphocyte count is greater than 115 cell/mm ³Patient in, the unmanned fatal event that takes place, even those are because the intermediate value CD on the baseline before the therapeutic alliance ₄Level has only 201, and immune system obviously worsens, and is the patient who is in the very big danger like this, also is like this.

With to put into practice the present invention consistent, after the basis of control virus replication is accomplished fluently, can add lymphokine advisably such as interleukin and interferon one class.

List of references

1. Larder，B.A.et al pp.436-441 Antimicrobial

Agents and Chemotherapy.Vol.34，March 1990.

2. Larder，B.A.et al.pp.1731-1734，Science，

Vol.243，1989.

3. Jackson，J.B.et al.pp.1416-1418.Journal

Clin.Microbiology，Vol.216，1990;Thompson，

J.D.et al.pp.371-378，Analytical Biochem.

Vol.182，1989.

4. Berenbaum，M.C.pp.269-333.Advances in Cancer

Research Vol.35，1981.

5. Fischl，M.et al.p.727-733，Annals of

Internal Medicine Vol.112，1990.

6.Pluda，J.M.et al pp.276-282

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Claims (13)

1, a kind of drug combination treatment system can prevent nucleoside analogue drug-resistant virus and/or improves the state of an illness by this therapeutic alliance, has the state of an illness relevant with immunocyte to the nucleoside drug-resistant virus especially in advance.

2, delay, reduce, or not only delay but also reduce the method that occurs nucleoside analogue drug-resistant virus in hiv infected patient, this method takes the wrong paired dsRNA of effective dose for said patient before being included in and using the treatment of nucleoside analog antiviral agents.

3, improve by the method for the peripheral blood mononuclear cell state of an illness of the HIA infected patient of nucleoside analogue drug-resistant, this method comprises the wrong paired dsRNA that takes effective dose to said patient.

4, the method for claim 2, wherein patient takes the nucleoside analog antiviral agent after using the paired dsRNA treatment of said mistake.

5, the method for claim 3, wherein said cell is the CD4 lymphocyte.

6, claim 2 or 3 method, the paired dsRNA of wherein said mistake is and the compound polyadenylic acid of polyuridylic acid.

7, the method for claim 6, wherein wrong paired dsRNA is the complex of polyinosine as poly-cytidine, contains 1/4 to 1/29 uracil or guanine base in the poly-cytidine.

8, the method for claim 7, wherein wrong paired dsRNA is rI _nR(C _{11-14 '}U) _nOr be the rI that contains the bond fission zone and show favourable treatment ratio character _nR(C _{11-14 '}U) _n

9, the method for claim 6, the wrong paired dsRNA amount of wherein taking can be so that should the paired dsRNA level of mistake reach 2 to 1000 μ g/ml in the circulation of patient's systemic blood.

10, claim 2 or 3 method, wherein dsRNA is the oligonucleotide of short chain, the general formula that defines this oligonucleotide is:

5 '-block-(I) _n-blockade 3 '

3 '-block-(C) _m-blockade 5 '

Wherein m and n are separately greater than 5, and less than 100, I is single phosphorylation inosine, and C is single phosphorylation cytidine, or

5 '-block-((I) _xA) _j-blockade 3 '

3 '-block-((C) _yU) _k-blockade 3 '

Wherein x and y be separately greater than 5, and less than 25, j and k are at least 1 and less than 10 separately, and the definition of I and C is the same, and A is not for being I and nucleotide, and U is base and the paired nucleotide of A, or

5 ' (I)-hinge-(C) _m3 '

N wherein, m, the definition of I and C is the same, and the blockade base complementation on a blockade base on the chain and the relative chain is provided.

11, according to the method for claim 10, wherein oligonucleotide stablize by the pairing of the inside of complementary non-homogeneous high polymer, and blockade base or hinge base or the two all contain the zone of complementary non-homogeneous high polymer.

12, the method for claim 2,3 or 4, wherein said nucleoside analog antiviral agent is Zidovudine, didanosine, zalcitabine, perhaps they and use medicine.

13, claim 2 or 3 method, the paired dsRNA of wherein said mistake is rI _n(C _{11-14 '}U) _n

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