ODN-1826 targets Toll-like receptor 9 (TLR9), an intracellular pattern recognition receptor that recognizes unmethylated CpG DNA motifs, activating innate immune responses and promoting Th1-polarized adaptive immunity.

In RAW 264.7 cells, ODN 1826 (1 μg/mL, 24 h) can increase NO and iNOS production [2].

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In RAW 264.7 cells, ODN-1826 (1 ug/mL, 24 h) stimulates NO and iNOS production. It induces apoptosis in immune cells and enhances immunostimulatory activity, serving as an antitumor agent and cardiac protector. The human counterpart sequence is ODN-2006.

In a model of chronic vascular damage, ODN 1826 (18 nM, subcutaneous injection, three times a week for seven weeks) increases the size of aortic atherosclerotic plaque [1]. In the Lewis lung cancer tumor model, ODN 1826 (0.05 mg, intraperitoneal injection, 1, 3, 5, 8, 11, 13 days) has combined anti-tumor growth effects [3].

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In murine tumor models (e.g., Lewis Lung Cancer), ODN-1826 (0.05 mg, i.p., on days 1,3,5,8,11,13) delays tumor growth, decreases tumor weight, and increases survival. In chronic vascular injury models, s.c. administration (18 nM, 3×/week for 7 weeks) increases aortic atherosclerotic plaque size compared to vehicle.

Not applicable, as ODN-1826 is an oligonucleotide agonist of the intracellular TLR9 receptor. Binding is assessed in cellular assays measuring TLR9 activation and downstream signaling. Standard receptor binding assays with purified TLR9 are not typically performed.

Western Blot Analysis[2]
Cell Types: RAW 264.7
Tested Concentrations: 1 μg/mL
Incubation Duration: 24 h
Experimental Results: Increased production of NO and iNOS.

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RAW 264.7 macrophages or other TLR9-expressing cells are seeded in 6- or 96-well plates. ODN-1826 (0.1-10 ug/mL) is added directly to the culture medium for 6-48 hours. After incubation, culture supernatants are collected for NO measurement (Griess assay), iNOS levels (Western blot or ELISA), and cytokine production (IL-6, TNF-alpha, IL-12) by ELISA. Apoptosis is assessed by flow cytometry using Annexin V/PI staining.

Animal/Disease Models: Lewis lung cancer mouse tumor model [3]
Doses: 0.05 mg (1, 3, 5, 8, 11, 13 days)
Route of Administration: intraperitoneal (ip) injection
Experimental Results: Delayed tumor growth, diminished tumor weight and increased Apoptotic tumor cells.

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In murine tumor models (e.g., Lewis Lung Cancer), ODN-1826 (0.05 mg/mouse) is administered intraperitoneally on days 1,3,5,8,11,13 post-tumor inoculation. Tumor volume is measured by calipers. Mice are euthanized and tumors are weighed. Serum and tumor tissue cytokine levels (IL-12, IFN-gamma) are measured by ELISA. Splenocyte activity is assessed by ex vivo stimulation.

After intravenous or subcutaneous administration in rodents, ODN-1826 is rapidly cleared from circulation (half-life of 15-60 min) but is taken up by immune cells in lymphoid tissues (spleen, lymph nodes). It accumulates in endosomal compartments of TLR9-expressing cells. Water solubility: 50 mg/mL (7.86 mM).

Standard oligonucleotide safety precautions apply. Systemic administration may induce pro-inflammatory cytokine release syndrome in sensitive individuals. Dose-dependent immunostimulatory effects observed. For research use only, not for human therapy. Avoid inhalation and contact with skin/eyes.

[1]. Proinflammatory Stimulation of Toll-Like Receptor 9 with High Dose CpG ODN 1826 Impairs Endothelial Regeneration and Promotes Atherosclerosis in Mice. PLoS One. 2016 Jan 11;11(1):e0146326.

[2]. CpG ODN activates NO and iNOS production in mouse macrophage cell line (RAW 264.7). Clin Exp Immunol. 2002 Jun;128(3):467-73.

[3]. CpG oligodeoxynucleotide 1826 enhances the Lewis lung cancer response to radiotherapy in murine tumor. Cancer Biother Radiopharm. 2011 Apr;26(2):203-8.

[4]. The toll-like receptor 9 agonist, CpG-oligodeoxynucleotide 1826, ameliorates cardiac dysfunction after trauma-hemorrhage. Shock. 2012 Aug;38(2):146-52.

This is a research-grade immunostimulatory oligonucleotide. It is the mouse-active CpG ODN corresponding to the human-active ODN-2006 (Agatolimod). It is not an approved drug but has been studied in preclinical and clinical vaccine adjuvants and cancer immunotherapy. Sequence: 5‘-tccatgacgttcctgacgtt-3'. MW ~6364 g/mol.

6138

202668-42-6

FITC-labeled ODN 1018 sodium;Biotin-labeled Agatolimod sodium;Biotin-labeled ODN 1826 sodium;FITC-labeled ODN 1826 sodium;Biotin-labeled ODN 1018 sodium

49771843

White to off-white solid powder

-40.5

46

145

118

406

18800

0

CC1=CN(C(=O)NC1=O)C2CC(C(O2)COP(=O)(O)OC3CC(OC3COP(=O)(O)OC4CC(OC4COP(=O)(O)OC5CC(OC5COP(=O)(O)OC6CC(OC6COP(=O)(O)OC7CC(OC7COP(=O)(O)OC8CC(OC8COP(=O)(O)OC9CC(OC9COP(=O)(O)OC1CC(OC1COP(=O)(O)OC1CC(OC1COP(=O)(O)OC1CC(OC1COP(=O)(O)OC1CC(OC1COP(=O)(O)OC1CC(OC1COP(=O)(O)OC1CC(OC1COP(=O)(O)OC1CC(OC1COP(=O)(O)OC1CC(OC1COP(=O)(O)OC1CC(OC1COP(=O)(O)OC1CC(OC1COP(=O)(O)OC1CC(OC1COP(=O)(O)OC1CC(OC1COP(=O)(O)O)N1C=C(C(=O)NC1=O)C)N1C=CC(=NC1=O)N)N1C=CC(=NC1=O)N)N1C=NC2=C(N=CN=C21)N)N1C=C(C(=O)NC1=O)C)N1C=NC2=C1N=C(NC2=O)N)N1C=NC2=C(N=CN=C21)N)N1C=CC(=NC1=O)N)N1C=NC2=C1N=C(NC2=O)N)N1C=C(C(=O)NC1=O)C)N1C=C(C(=O)NC1=O)C)N1C=CC(=NC1=O)N)N1C=CC(=NC1=O)N)N1C=C(C(=O)NC1=O)C)N1C=NC2=C1N=C(NC2=O)N)N1C=NC2=C(N=CN=C21)N)N1C=CC(=NC1=O)N)N1C=NC2=C1N=C(NC2=O)N)N1C=C(C(=O)NC1=O)C)O

VQWNELVFHZRFIB-UHFFFAOYSA-N

InChI=1S/C194H249N67O125P20/c1-77-40-248(188(280)235-167(77)263)130-20-84(262)104(348-130)47-329-388(290,291)375-92-28-138(250-42-79(3)169(265)237-190(250)282)356-112(92)55-339-403(320,321)383-100-36-146(258-73-217-153-163(258)227-178(204)231-174(153)270)364-120(100)63-344-393(300,301)372-89-25-135(246-18-12-128(199)225-186(246)278)353-110(89)53-334-400(314,315)380-97-33-143(255-70-214-150-157(201)208-67-211-160(150)255)362-118(97)61-342-405(324,325)385-102-38-148(260-75-219-155-165(260)229-180(206)233-176(155)272)366-122(102)65-346-398(310,311)378-95-31-141(253-45-82(6)172(268)240-193(253)285)357-113(95)56-336-391(296,297)370-87-23-133(244-16-10-126(197)223-184(244)276)349-106(87)49-330-390(294,295)369-86-22-132(243-15-9-125(196)222-183(243)275)352-109(86)52-333-396(306,307)376-93-29-139(251-43-80(4)170(266)238-191(251)283)358-114(93)57-337-397(308,309)377-94-30-140(252-44-81(5)171(267)239-192(252)284)360-116(94)59-340-404(322,323)384-101-37-147(259-74-218-154-164(259)228-179(205)232-175(154)271)365-121(101)64-345-394(302,303)373-90-26-136(247-19-13-129(200)226-187(247)279)354-111(90)54-335-401(316,317)381-98-34-144(256-71-215-151-158(202)209-68-212-161(151)256)363-119(98)62-343-406(326,327)386-103-39-149(261-76-220-156-166(261)230-181(207)234-177(156)273)367-123(103)66-347-399(312,313)379-96-32-142(254-46-83(7)173(269)241-194(254)286)359-115(96)58-338-402(318,319)382-99-35-145(257-72-216-152-159(203)210-69-213-162(152)257)361-117(99)60-341-392(298,299)371-88-24-134(245-17-11-127(198)224-185(245)277)350-107(88)50-331-389(292,293)368-85-21-131(242-14-8-124(195)221-182(242)274)351-108(85)51-332-395(304,305)374-91-27-137(355-105(91)48-328-387(287,288)289)249-41-78(2)168(264)236-189(249)281/h8-19,40-46,67-76,84-123,130-149,262H,20-39,47-66H2,1-7H3,(H,290,291)(H,292,293)(H,294,295)(H,296,297)(H,298,299)(H,300,301)(H,302,303)(H,304,305)(H,306,307)(H,308,309)(H,310,311)(H,312,313)(H,314,315)(H,316,317)(H,318,319)(H,320,321)(H,322,323)(H,324,325)(H,326,327)(H2,195,221,274)(H2,196,222,275)(H2,197,223,276)(H2,198,224,277)(H2,199,225,278)(H2,200,226,279)(H2,201,208,211)(H2,202,209,212)(H2,203,210,213)(H,235,263,280)(H,236,264,281)(H,237,265,282)(H,238,266,283)(H,239,267,284)(H,240,268,285)(H,241,269,286)(H2,287,288,289)(H3,204,227,231,270)(H3,205,228,232,271)(H3,206,229,233,272)(H3,207,230,234,273)

[5-(2-amino-6-oxo-1H-purin-9-yl)-2-[[[2-[[[2-[[[5-(2-amino-6-oxo-1H-purin-9-yl)-2-[[[2-[[[2-[[[2-[[[2-[[[2-[[[5-(2-amino-6-oxo-1H-purin-9-yl)-2-[[[2-[[[2-[[[5-(2-amino-6-oxo-1H-purin-9-yl)-2-[[[2-[[[2-[[[5-(4-amino-2-oxopyrimidin-1-yl)-2-[[[5-(4-amino-2-oxopyrimidin-1-yl)-2-[[hydroxy-[5-(5-methyl-2,4-dioxopyrimidin-1-yl)-2-(phosphonooxymethyl)oxolan-3-yl]oxyphosphoryl]oxymethyl]oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]-5-(6-aminopurin-9-yl)oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]-5-(6-aminopurin-9-yl)oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]-5-(6-aminopurin-9-yl)oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-3-yl]oxy-hydroxyphosphoryl]oxymethyl]oxolan-3-yl] [3-[hydroxy-[[3-hydroxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy]phosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methyl hydrogen phosphate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~50 mg/mL (~7.86 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)