JPS61161204A - Anti-plant-viral - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application There are many plant virus diseases, many of which are the most contagious types. Statistics show that these diseases cause millions of dollars in damage every year. Such viral diseases are particularly difficult to control and virtually impossible to reverse. A single plant virus can attack many different plant species and can attack many different vectors (insects, nematodes,
molds, parasitic vascular plants, and agricultural tools). Viruses can be transmitted from one generation to another through seeds, and the eelfish virus is born in the soil.

In mammals, interferon is the most rapid defense mechanism against viral infection. One activity induced by interferon in mammalian cells is AT.
By converting P into a series of unique 2',5'-oligoadenylates, the best power, varnish [edl(
+981>Methods in Enzymology = Volume 78 and 7
Volume 9, Academic Press, New York). Similar but non-identical plant oligonucleotides are introduced into tobacco plants by AVF (antiviral factor or plant interferon) after Taba comosaic virus (TMV) infection (Debash, Wy et al. (1981) ) Pyroroyo Ill: 103-112)
.

Recently, natural 2',5'-diphospho-guanine (A3'p
p5'-oligoadenylate as well as chemically synthesized 2',5'-diphospho-guanine (A3'pp5'-oligonucleotide analogs) potently inhibit TMV replication at nanomolar (10-9 M) levels. It has been reported that it is an agent. (Tashiko, Y., Hietzkoos, S., and T.Jin. (1982) Science 216
: 1415-1416;
84) Jour, Biol.

Chew, 259:3482-3488).

Means for Solving the Problem The present invention relates to the use of nucleotides conjugated by pyrophosphoryl linkages as anti-plant viral agents. Particularly exemplified are dinucleotides 5', 5'-P', p2 to block TMV replication in tobacco plants when applied through the leaves or roots of tobacco plants.
-diphosphate, diadenosine 21°5'-p1,p
2-diphosphate and diadenosine 3', 5'-1
1'+p2-diphosphate.

Application of these antiviral compounds through the roots of tobacco plants or on the leaves of the plants results in inhibition of TMV replication.

The military nucleotides of the invention that can be used to inhibit plant virus replication are known or available compounds or compounds that can be readily prepared by procedures well known in the art. These conjugated dinucleotides can be represented by the general formulas I to II of Char) A, where X1 is A or C or 0 or E, and X2 and X3 are B or C or D or E. where R1 and R2 are purines or pyrimidines or analogs thereof, and R3, R4, R5, R6, R7, R8, R9, and RIG are H1OR',5'-diphospho-guanine (A3
'ppNR''R'' (wherein R' and R'' are the same or different and are ■ or alkyl of 1 to 4 carbon atoms), amino acid, θ-COR', 5'-diphospho-
Guanine (A3'pp-COR", halogen, C0OR
”, -504, or -po4, and R11 is P or P
O(-) or s=.

or S, R12 is 0 or S, R+3 and R
IB is 0, S, C112 or Se, R14 and R15 are 0 or C112, R17, R18, R
19・and R20 are 11・011・N 112 C
It a and R21 and R22 are Hl 011. N
11COCH3, and n in ■, ■, or ■ is an integer 2 to 3
and n' is an integer from 4 to 16.

Examples of purines and pyrimidines are adenine, guanine, cytosine, uracil, thymine.

Examples of purine and pyrimidine analogs are 6-dimethylaminopurine, 6-medoxypurine, 8-azaadenine, 2-
Chloropurine, 5-azacytosine, sangivamycin,
Aglycones of toyocamycin, formycin and tshercisin, 8-bromopurine and 1. N6-nitenoadenine. These purine and pyrimidine nucleotides are commercially available (Sigma Chemical Company, MI-H St. Louis). Can be chemically condensed to pyrophosborylated anti-plant virus dinucleotides using the methods disclosed herein.

Halogens are chlorine, bromine, and fluorine.

The amino acid can be any one of the 20 D- or L-common amino acids, namely alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine. , proline, serine, threonine, tryptophan, tyrosine and valine.

The following nucleoside molecules include triphosphates, which are suitable precursors for the synthesis of birophosphorylated anti-plant viral nucleotides within formula I, such as adenosine, 21- or 3 '-deoxyadenosine, 2'- or 3'-acetamido-β-D-ribofuranosyl adenine, 2'- or 3'-amino-3'-deoxy-β-D-ribofuranosylcytosine, l-β- D-xylofuranosylguanine, and 1-β-D-arahynofuranosyluridine. These nucleotides are commercially available (Sigma) and chemically dinucleotides can be prepared using the methods disclosed herein or other well-known methods.
' if, 5'-p1, pn (m=2'' or 3'; n=
2-3) Can be condensed to n-phosphate.

Precursors for the organic synthesis of antiviral compounds, ie nucleotide 21- or 3'- or 5'-monophosphates, are commercially available (Sigma).

The integer n is the 2' of the dinucleotide, the 3' of the 5'-sentence,
5'- or 5', 5'-diester repeating internal bond.

Pyrophosphoryl Compounded Anti-Plant Virus Dinucleotides (1965) J, Org, Chem
30, 3381-3387;
1978) Proc, Natl, Acad, Sci, 1
JSA 76 6081-6085; Kosa Rich
(1973) Biochemistry 124458-4463: *7D.

Amer, Chem, Soc, 87:17
85-1788; Nejku et al. (1985) J.

Med, Chem, 28 171-177; Teheras et al. (1985) J, Med, Chem.

284B-46; can be chemically prepared essentially as described in Zurak et al. (+985) Nucleosides and Nucleotides II:165-167. Using the chemical synthesis described above, a wide range of phosphorylated dinucleotides 11'+P"(n"2-3)n-phosphates can be made that differ in the nucleotide and its mono-, di-, and triphosphate moieties. Examples of such compounds are as follows.

Diadenosine 5',5'-diphosphoguanine (A3'
pp5″′-1′1,p2-diphosphate (Ap2A
), Diadenosine 5', 5'''-p'+p3-triphosphate (Ap3A), Diadenosine 5', 5
'j l-pl, p4-tetraphosphate (AI]4A), diadenosine 5''.

5″′-F”+p5~pentaphosphate (AI) 5A
), and diadenosine 51.5″′-1″++″-hexaphosphate (ApeA) (all available from Sigma) and adenosine 5′,5′-diphospho-guanine (A3′pp5″′-p′+p2 , 5'-diphospho-guanosine (Al12G), adenosine 5Z5'''-1''
+92,5'-diphospho-uridine (Ap2U), adenosine 5' + 5'''-1'' + 1''-diphosphocytidine (A112 f': ), 2',3
”-cyclic phosniadenosine 5′.

511+-p1,p2-diphosphate (p<Ap2A
>p), guanosine [5']) lyphosphate adenosine (GρaA), thymidine [5' cosyphosphate adenosine (TI] 2A), cytosine [5'] tetraphosphate cytosine (CI) 4C), adenosine 3'', 5′
-diphosphoadenosine (A3'pp5''A), adenosine 3',5'-diphosphocytidine (A3'1)l)
5'C), adenosine 3', 5',5'-diphospho-
Guanosine (A3'pf15'G), adenosine 2'
, 5'-diphosphoadenosine (A2'pp5'A),
Adenosine 2', 5'-diphosphocytidine (A2'
pp5″C), adenosine 2′,5′-diphospho-guanine (A3′pp5′,5′-diphospho-guanosine (
A2'pp5'G) etc.

The most preferred compound is diadenosine 5+, 5+++-1
1'++12-diphosphate (Ap2A), adenosine 5'.

5″′, p1, pl, 5′-diphospho-guanosine (
A112G), adenosine 5'+5'''-p'+p2,
5'-diphospho-uridine (Ap2U), adenosine 5
',5'''-p1,pl-diphosphocytidine (Al1
2C), 2',5'-diphospho-guanine (A3'pp
3'-cyclic bosphodiadenosine 5',5'''-p1,
pl-diphosphate (Il<Ap2A>+1), and adenosine 5',5'-diphospho-guanine (A3'p
p5'-diphospho-thymidine.

The second most preferred compound is adenosine 2',5'-diphospho-guanine (A3'pp5'-diphosphoadenosine (A2'pp5'A), adenosine 2+.

5',5'-diphospho-guanosine (A2'1ll15
'G), adenosine 2', 5'-diphospho-uridine (A2'pp5'U), adenosine 2',5'-diphospho-guanine (A3'pp5'-diphosphocytidine (
A2'pp5'C) and adenosine 2',5'-diphosphothymidine 2'pp5'T).

The third most preferred compound is adenosine 3',5'-diphospho-adenosine (A3'pp5'A), adenosine 3',5',5'-diphospho-guanosine (A3'p
p5'G), adenosine 3',5',5'-diphospho-
uridine (A3'pp5'tJ), adenosine 3',
5'-diphosphocytidine (A3'pp5'C) and adenosine 3'',5'-diphosphothymidine (A3'pp5'
T).

All of the above compounds are from P.M., Milwaukee, Wyoming.
-L Biochemicals.

Application of anti-plant virus replication effective amounts of phosphorylated dinucleotides to plants susceptible to plant virus infection allows the treated plants to resist virus attack. Although the method of the invention is exemplified herein by the use of tobacco plants and TMV, the method of the invention can be applied to a variety of plants,
For example, tobacco, barley, beans, oats, beets, soybeans,
It can be used to treat sugar beets, sugar cane, wheat, alfalfa, cherries, citrus, grapes, melons, cucumbers, lettuce, pepper, potatoes, cacao, coconut, etc., and against a wide range of viruses, such as tobacco. Mosaic, Vein Banding, Strive Mosaic, Yellow Doula Oats, Yellow Mosaic, Common Mosaic,
Carlito Whelk, Soilbone Mosaic, Bud Blight, Streak Mosaic, Leaf Roll, Beanbot Molle Virus, Tobacco Ringspot Virus, Beannut Stand Virus, Peanut Mozzle Virus, Maize Dwarf Mosaic Complex, Soybean Mosaic Virus It can be used for viruses susceptible to , alfalfa mosaic virus, cucumber mosaic virus, tomato ringspot virus, etc.

Effective anti-plant virus replication amounts of phosphorylated dinucleotides can be readily determined for different plants and viruses using procedures disclosed herein or known in the art. This measurement can be easily made without extensive experimentation. The following is a disclosure of the materials and methods used in the present invention.

1. JL product Nicotiana glutinosa (Nicottana glu
t:nosa> and N, tabacum var, Samsam (N, tabacum var.

samsun) plants were grown in potting medium under greenhouse conditions (25±3° C. and constant lighting). Plants every two weeks -21-.

were given commercially available fertilizer. When plants are used at the 5-8 leaf stage, Debash Y9, Pigges Varnish, and Sera Eye, (19B2) Sciens 216: 1415-14
16; Debash Rai 0 etc. (1984) Jour,
Biol, Chem.

259:3482-3486).

2. TMV・9 - of
Assays for antiviral activity in leaf discs infected with TMV were conducted to determine whether TMV had spread systemically.
tabacum pal, samsan (N, tabacum v
ar, samsun). Leaves were mechanically washed with pure TMV (5% in sodium phosphate buffer 0.01M).
μgl...1, pH 7.6, containing 0.1 g/ml of carborundum as an abrasion agent) was inoculated.

Immediately after infection, leaf discs (6.5 mm diameter) were punched out from the inoculated leaves and placed in a beaker containing 0.01 M sodium phosphate at pHr, 6.

Groups of 20 discs were randomly drawn from this common pool and placed in separate Petri dishes, each containing 18 ml of 10 m M 4-(2-hydroxyethyl)-1. -piperazineethanesulfonic acid (HEP)
ES)-KOHpH7,05188mg/I Na2
It contained HPOn 7820. Discs are treated with antiviral dinucleotides for 1 hour by adding the compound with 2 ml of 600 mM CaCl2. 1
At the end of the hour incubation, the buffer containing antiviral nucleotides is replaced with a buffer without antiviral dinucleotides for an additional hour. Disk next 0.01M
Wash with sodium phosphate p) 17.6 and incubate for a further 72 hours in phosphate buffer (Debash Y9, Pigges Nys, & Sera I, (1982) Science 2
16:1415-1416. , Dabash Wai et al. (1
984) J. Biol. Chem.

259:3482-3486). The discs were homogenized in 0.01 molar sodium phosphate buffer pH 7.6 and TM
V content is determined by the enzyme-linked immunosorbent assay (TLISA) described below.

3. Weighing quantity
The activity of antiviral dinucleotides to block TMV replication in LtLflll intact N, Curtinosa plants (containing the N gene for TMV localization) was determined by infectivity tests. A solution containing 5 μg/ml TMV, 0.1 μg/ml carborundum and antiviral dinucleotides was applied to five half leaves of N. glutinosa.

The opposite half of the leaf served as a control and was inoculated with an infection solution containing TMV and carborundum, but no dinucleotide. Infection was allowed to stand for 488 hours. Inhibition of TMV replication was calculated as the % of localized disease developed in the half leaf treated with antiviral compounds compared to the control half leaf.

4, N, f of the brush of Tahanim
Virus Nicotiana tabacum (Ni) of c.
cotiana tabacum) plants were used as a source of plant tissue in which TMV was spread throughout. These plants did not contain the N-gene for TMV localization.

(a) Application to roots: Remove tabacum plants from their pots.
Transferred into a 501 tube containing 0 ml IO + nM HEPES-OH pH 7,6 and antiviral dinucleotide.

After 2 hours the plants were transferred to 50 ml tubes without antiviral compounds and inoculated with TMV as above. After 24 hours, the leaves were homogenized in 0.01 M sodium phosphate buffer pH 7.6 (Ig leaves/3 ml buffer) and TMV content was determined by ELISA.

(b) Application to leaves Antiviral activity was measured as described above, but N.

I used the tabacum plant. After 24 hours, half the leaves were removed, homogenized, and TMV content was determined by ELISA.

The TMV content of the LISA homogenate was essentially determined by Clark and Adams (
(+977) J, Gen, Virol, 34:475
-483), but the following microtiter (microt
iter) using the layer order on the plate.
Measured by.

(a) γ- of serum obtained from chickens immunized against TMV
The globulin fraction was absorbed directly onto the plate.

(b) The next layer was a test sample (various dilutions) or a series of purified TMV suspensions at known concentrations (standard curve).

(C) The third layer was γ-globulin from TMV-immunized rabbit serum.

(d) The last layer was goat antibody against rabbit immunoglobulin G conjugated with alkaline phosphatase (Sigma). Phosphate release from p-nitrophenyl phosphate for antibody-conjugated alkaline phosphatase was monitored spectrophotometrically using a microplate reader (Dynatec Diagnostics Inc., South Windham ME), which was approximately 2
It was proportional to the degree of TMViI in a log-linear manner. (<2 ng ~> 200 ng) Debash Y 1,
[1982] Science 216:1415-1416) 6, M. γ- of serum obtained from chickens and rabbits immunized against TMV.
The globulin fraction was a generous gift from Professor Iran Sera of the Department of Agriculture, Hebrew University of Jerusalem, Rehopot, Israel.

The following example describes procedures, including the best mode, for carrying out the invention. These examples are not to be construed as limiting.

1LnJLl -9 1 Bako's -26 Isshin's 5'5'-1nn=2-3-"shi゛"+5'-p'pn (n=2-3)-dinucleotide in TMV-infected disks It was as effective as 2',5''-adenylate trimer nuclei in blocking TMV replication (Table 1). As demonstrated, the calcium phosphate co-precipitation technique Thank you for your cooperation (synergization).

However, the calcium co-precipitation technique was only required in the disc assay and not in the other inhibitory activity measurements disclosed above. The results in Table 1 show that the calcium co-precipitation technique enhances antiviral compounds by increasing their uptake in the disk assay. Table 1 also demonstrates the importance of calcium coprecipitation in synergizing antiviral dinucleotides in the TMV-infected leaf disc assay. This result indicates that the coprecipitation technique increases the uptake of antiviral dinucleotides.

5' containing n=4 or more initial phosphates
, 5°-p'p' (n=2-3) dinucleotides had less antiviral activity compared to dinucleotides with 2 and 3 internal phosphates. However, modification of the nucleotide moiety (i.e. one of the adenosines is changed to G, C
, U, addition of external phosphates or addition of external 2',3'-cyclic phosphates (e.g. p<A5'pp5'A>ρ) did not reduce their antiviral potency. Ta.

table".

[Ca”2] Co-precipitation [Ca+2 Co-precipitation]
L-No "LQ" L-]00nM 200nM 1100n 200n
M control 75';Ob 75;0 56;0
56;02',5'-diphosphoguanine (A3'pp
5'ApApA 46:3936:52 53;5
48;14A5'pp5'A 48;36 44;
41 N, 1. :ON, 1. :OA5'ppρ5'A
65; 13 38; 50 N, 1. ;ON, 1.
; Oa = ngTMV/m lb = % inhibition relative to control N, 1. = 5',5' to suppress TMV replication in intact plants without inhibition
The ability of -diphospho-guanine (A3'pp5'-dinucleotides) was tested using N. glutinosa plants. 5',5'-dinucleotides inhibit the formation of local lesions after infection (Table 2).

The most interesting is A5'ppASA5'ppp5'A,
1 nM (1
Application at a concentration of 0-6 M) resulted in induction of resistance to TMV infection even in the untreated half of the leaves.

The number of local diseases on the untreated half leaves was reduced compared to the control. Therefore, antiviral compounds induced antiviral resistance in plant tissues even without direct application.

Table 2 10nM (to-8M) 100nM (to-71
00n le
几2',5'-diphosphoguanine (A
3'pp5'-ApApA 8B, 8 48.6
B9.4 9.8A5'pp5'A 96
．． 0 68.4 95.4 54.6A5'ppp
5'A 9B, 6 80.3 100 100
A5'pp5'G 84.8 59.8 8
9.5 44.7A5'pp5'C55,036,15
',935,05'5'lJ 38.1 5.
0 01000nM (101M)2',
5'-diphospho-guanine (A3'pp5'-ApAp
A 100 100A5'pp5'A 10
0 100A5'''')p5'A 100 1
00A5'pρ5'G 100 10 0A5'pp5'C63,532,95"U
O,,0 Note: Results are % protection of TMV infected leaves compared to control.

=30- Example 3 5'5'-11n=2~3''
of the induced resistance of N- for virus localization.
A comparison was made between foliar and root application of antiviral dinucleotides to N, Tahakum plants to establish a relationship to gene and compound delivery capacity. N, tabacum plant 10m1]OmM+1EPEs
- in a vial containing KOJI pH 7, 6 and 1μ antiviral dinucleotide. Plant roots were left in the solution for 2 hours and then transferred to vials containing buffer without antiviral compounds.

The plants were then infected with TMV as above (see section 4 (page 22), Antiviral activity of N. tabacum on intact plants, paragraph (a)). Another N, Barra Bluff (
Tested as described in b).

1 μM (10−8 M) of 2′,5′-diphospho-guanine (A3′pp5′-ApApA, A5′pp5′A
Exposure of plant roots to A5'pp5'G, or A5'pp5'G resulted in total inhibition (98-100% inhibition) of the virus (
Table 3).

The induced resistance in leaf areas not directly treated with antiviral dinucleotides cannot be attributed to the N-gene since resistance was also observed in N. tabacum plants.

- Side control 2000 1 μM 2',5'-diphospho-guanine (A3'pp
5'ApApA O+001μM A5'pp5'
A O1100Iu A5'ppp5'A
98 51 T in sexually transmitted TMV-infected disks
31.5l-plp2-dinucleotide inhibits MV replication by 2',5'-diphospho-guanine (A3'p2-dinucleotide).
It was as effective as the p5'-adenylate trimer core. (Table 2) Suppression of TMV replication in TMV-infected leaf discs by p2-diadenosine birophosphate and 2',5'-ApApA9 〇 T v/m1
Comparison of text and edict 5500 2', 5'ApApA 2xlO1M 200 B4 20xlO-9M +80 67200xlO-
9M too 82A3'pp5'A 2xlO-9M 550 O2Oxl0-9
M 200 6400x OlM +50
73 The results in Table 4 show that A3''pp5'A and 2+ at equivalent concentrations.

5'ApApA has been demonstrated to confer similar antiviral status.

The ability of A3'pp5'A to suppress TMV replication in intact plants was tested using N. glutinosa plants. A3'pp5'A suppresses TMV replication as shown by inhibiting local lesion formation after infection. Dabash Kui et al. (1984) J, Biol, Ch.
Em, 259:3482-3486). Application of A3'pp5'A at a concentration of 1μ in the infection solution resulted in the introduction of resistance to TMV infection even in the untreated half of the leaves (Table 5).

Table j.

Control 00 10-6M 2',5'-diphospho-guanine (A3'
pp5'-ApApA90to.

O-6MA'5' 93 100
Note: Results above are % protection compared to control TMV infected leaves.

□152-1? -2'+5'-p1p'-diadenosine was effective in suppressing TMV replication in single TMV-infected disks (
Table 6).

Table 6 Control □b Adenosine 2',5'-diphospho-guanine (A3'p
p5'-1"+p2-diadenosine 10-10 MO 10-9M 21 10'-"M 26.410-7
M 37 1-6M 42 b=% inhibition relative to control The above results are (1) m=5'-1)'II) n-(m=
2',5'-diphospho-guanine (A3'pp3',5
'; n = 2-3) - antiviral ability of dinucleotides, (2) advantages of root application (application through irrigation)
This is due to active transport of phosphate-containing compounds, and (3""""-1]'+
I"-(11"2', 3', 5"; n=2-3)
- the property of the dinucleotide to induce resistance to viral infection even in plant areas not directly treated;
(due to active rabbit transport or triggering of an antiviral state).

It is noteworthy that application of antiviral compounds in the range of 10-9 M to 10-6 M resulted in total suppression of the virus without any apparent phytotoxicity.

Chart A E Chart A (2nd sheet) 11th chart A (3rd meter) C Chart A (4th sheet) [CH3 (CH2)n, CH2-] ]n'=
4-1-40-

Claims (1)

[Scope of Claims] 1. 5', 5'-;2',5'-; or 3' anti-plant virus replication amount effective for plants susceptible to plant virus infection;
, 5'-dinucleotide. 2. The method according to claim 1, wherein the susceptible plant is a tobacco plant and the plant virus is tobacco mosaic virus. 3. The above 5', 5'-;2',5'-; or 3', 5'
2. A method according to claim 1, wherein the dinucleotide is applied to said susceptible plants on the leaves or through the root system. 4. Plants susceptible to the above plant viruses are treated with an anti-plant virus replication effective amount of 5',5'-dinucleotide (formula I);
2',5'-dinucleotide (formula II) or 3',5'-
A method of inhibiting plant replication in susceptible plants consisting of treatment with dinucleotides (formula III) ▲ Mathematical formulas, chemical formulas, tables, etc. There are chemical formulas, tables, etc.▼III In the formula, X_1 is A, C, D, or E, and n is an integer 2
~3, where X_2 and X_3 are B, C, D, or E, and n is an integer 2 to 3, A is the formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼, and B is the formula ▲ There are mathematical formulas, chemical formulas, tables, etc.▼, and C is a formula▲There are mathematical formulas, chemical formulas, tables, etc.▼, and D is [CH_3(CH_2)_n' H_2-], where n' is an integer. 4 to 16, E is ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ In the formula, R_1 and R_2 are purines or pyrimidines or analogs thereof, and R_3, R_4, R_5, R
_6, R_7, R_8, R_9, and R_1_0 are H,
OR', NR'R'' (wherein R' and R'' are the same or different and are H or alkyl of 1 to 4 carbon atoms), amino acid, -OCOR', -COR', halogen , COOR'', -SO_4, or -PO_4, R_1_1 is P or PO(-) or S=O or S, R_1_2 is O or S, R_1_3 and R_
1_6 is O, S, CH_2 or Se, R_1_4
and R_1_5 is O or CH_2, R_1_7,
R_1_8, R_1_9, and R_2_0 are H, OH,
NH_2CH_3 and R_2_1 and R_2_2 are H
, OH, NHCOCH_3. 5. In the method of inhibiting plant virus replication in plants susceptible to plant viruses, the above plants are treated with the formula ▲ mathematical formula, chemical formula,
There are tables etc. ▼ 5' of [In the formula, R1 and R2 are purines or pyrimidines or their analogs, and n is an integer 2 or 3]
5. The method of claim 4, which comprises treating the plant with an effective amount of -,5'-dinucleotide to inhibit plant virus replication. 6. The above 5',5'-nucleotide is diadenosine-5
′,5″′-p^1,p^2-diphosphate (Ap_
2A), adenosine-5',5'''-p^1, p^2-
Diphosphoguanosine (Ap_2G), adenosine-5'
,5″′-p^1,p^2-diphosphouridine (Ap_
2U), adenosine-5',5″'-p^1, p^2-
Diphosphocytosine (Ap_2C), 2',3'-cyclic phosphodiadenosine, 5',5'''-p^1,p^2-diphosphate, (p<Ap_2A>p) and adenosine 5'5'- 7. The method according to claim 1, wherein the 2',5'-dinucleotide is adenosine 2'.
, 5'-diphospho-adenosine (A2'pp5'A),
Adenosine 2',5'-diphospho-guanosine (A2'
pp5'G), adenosine 2',5'-diphospho-uridine (A2'pp5'U), adenosine 2',5'-diphospho-cytidine (A2'pp5'C) and adenosine 2',5'-diphospho-uridine Thymidine (A2'pp5'T)
The method according to claim 4. 8. The above 3',5'-dinucleotide is adenosine 3'
, 5'-diphospho-adenosine (A3'pp5'A),
Adenosine 3',5'-diphosphoguanine (A3'p
p5'G), adenosine 3',5'-diphospho-uridine (A3'pp5'U), adenosine 3',5'-diphospho-cytidine (A3'pp5'C) or adenosine 3
5. The method according to claim 4, wherein the compound is ',5'-diphospho-thymidine (A3'pp5'T). 9,5',5'-dinucleotide (formula I) ▲There are mathematical formulas, chemical formulas, tables, etc.▼ I or 2',5'-dinucleotide (formula II) ▲There are mathematical formulas, chemical formulas, tables, etc.▼II or 3',5'-Dinucleotide (Formula III) ▲Mathematical formulas, chemical formulas, tables, etc.▼III [In the formula, X_1 is A or C or D or E, n is an integer 2 to 3, and X_3 is B or C or D or E, n is an integer 2 or 3, and n is an integer A is a formula ▲ There are formulas, chemical formulas, tables, etc. ▼ B is a formula ▲ Mathematical formula, chemical formula , tables, etc. ▼ , and C is a formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ , and D is [CH_3(CH_2)_n'CH_2-], where n' is an integer from 4 to 16. , E is ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ In the formula, R_1 and R_2 are purines or pyrimidines or analogs thereof, and R_3, R_4, R_5, R
_6, R_7, R_8, R_9, and R_1_0 are H,
OR', NR'R'' (wherein R' and R'' are the same or different and are H or alkyl of 1 to 4 carbon atoms), amino acid, OCOR', -COR', halogen, COOR'', -SO_4, or -PO_4,
R_1_1 is P or PO(-) is S=O or S,
R_1_2 is O or S, R_1_3 and R_1_
6 is O, S, CH_2 or Se, R_1_4 and R_1_5 are O or CH_2, R_1_7, R_
1_8, R_1_9, and R_2_0 are H, OH, NH
_2CH_3, R_2_1 and R_2_2 are H, O
H, NHCOCH_3]. 10. 5' of the formula ▲ Numerical formula, chemical formula, table, etc.
10. The composition of claim 9, comprising a -,5'-dinucleotide. 11. The above 5',5'-nucleotide is diadenosine-
5′,5″′-p^1, p^2-diphosphate (Ap
_2A), adenosine-5',5'''-p^1, p^2
-diphospho-guanosine (Ap_2G), adenosine-
5',5'''-p^1, p^2-diphospho-uridine (
Ap_2U), adenosine-5′,5″′-p^1,p
2-diphospho-cytidine (Ap_2C), 2',3'-
Cyclic phosphodiadenosine 5',5'''-p^1, p^2
-diphosphate, (p<Ap_2A>p) and adenosine 5'5'-diphospho-thymidine (Ap_2T). 12. The above 2',5'-dinucleotide is adenosine 2
',5'-diphospho-adenosine (A2'pp5'A)
, adenosine 2',5'-diphospho-guanosine (A2
'pp5'G), adenosine 2',5'-diphospho-uridine (A2'pp5'U), adenosine 2',5'-
Diphospho-cytidine (A2'pp5'C) and adenosine 2',5'-diphospho-thymidine (A2'pp5'T)
) The composition according to claim 9. 13. The above 3',5'-dinucleotide is adenosine 3
',5'-diphospho-adenosine (A3'pp5'A)
, adenosine 3',5'-diphosphoguanine (A3'
pp5'G), adenosine 3',5'-diphospho-uridine (A3'pp5'U), adenosine 3',5'-diphospho-cytidine (A3'pp5'C) and adenosine 3',5'-diphospho-uridine Thymidine (A3'pp5'T)
The composition according to claim 9.