HCMV(IC50= 0.34 μM);HCMV(MCMV=0.039 μM)
HCMV (EC50 ranged from 0.15 to 1.97 μM across laboratory strains and clinical isolates; specific EC50 values: Davis 0.52±0.09 μM, AD169 0.52±0.14 μM, Towne 1.03±0.19 μM, clinical isolates He 0.89±0.10 μM, R.R. 1.43±0.42 μM, B.K. 1.97±0.52 μM, M.F. 1.50±0.25 μM, ganciclovir-resistant Ulm B 1.48±0.30 μM, ganciclovir/foscarnet double-resistant V1 0.15 μM, V2 0.30 μM, ganciclovir/cidofovir double-resistant isolate 2 1.50 μM) [2]
MCMV strain Smith (EC50 = 0.05 μM in plaque assay, 0.04 μM in cytopathogenicity assay) [2]

Strong anti-CMV agent tomeglovir (BAY 38-4766) has IC50 values for HCMV and MCMV of 0.34 μM and 0.039 μM, respectively. Additionally, HELF and NIH 3T3 cells are suppressed by tomeglovir, with CC50s of 85 μM and 62.5 μM, respectively[1]. At EC50s of 1.03 ± 0.57 μM and less than 1 μM, tomeglovir (BAY 38-4766) inhibits the HCMV Davis and different strains of monkey CMV[2].

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Tomeglovir showed antiviral activity against HCMV laboratory strains (Davis, AD169, Towne) and clinical isolates including ganciclovir-resistant, ganciclovir/foscarnet double-resistant, and ganciclovir/cidofovir double-resistant strains, with EC50 values ranging from 0.15 to 1.97 μM in plaque inhibition assays. The compound was not active against HSV-1, HSV-2, VZV, HHV-6, HIV, HBV, adenovirus type 5, or measles virus. It was active against monkey CMV strains and rodent CMV strains including MCMV (EC50 = 0.05 μM). [2]
Mechanistic studies: Southern blot analysis showed that Tomeglovir did not inhibit viral DNA synthesis (unlike ganciclovir), but specifically blocked cleavage of concatemeric DNA into monomeric genome-length fragments in a dose-dependent manner, with complete inhibition at ≥0.5 μM. [2]
Sequence analysis of resistant HCMV AD169 revealed mutations in UL89 and UL104; resistant MCMV Smith also had mutations in the murine homologues of UL89 and UL104. No cross-resistance to ganciclovir, cidofovir, or foscarnet was observed (resistance index ~1 for these drugs). [2]

Tomeglovir (BAY 38-4766; 3, 10, 30, 100 mg/kg, p.o.) decreases MCMV-DNA in the salivary glands, livers, and kidneys of NOD-SCID mice infected with MCMV in a dose-dependent manner while also extending the mice's survival. In the hollow fiber mouse model, tomeglovir (10, 25 and 50 mg/kg) exhibits antiviral activity[1]. In SCID mice infected with MCMV, tomeglovir (BAY 38-4766) exhibits antiviral activity; in rats and mice, the LD50 is greater than 2000 mg/kg[2].
In a lethal MCMV infection model in NOD-SCID mice (i.p. infection with 5×10^5 pfu MCMV Smith), oral administration of Tomeglovir twice daily for 8 consecutive days at doses of 3, 10, 30, and 100 mg/kg dose-dependently improved survival compared to placebo. Significance was observed at all dosages (P=0.02223 at 3 mg/kg, P=0.00375 at 100 mg/kg, log-rank analysis). [2]
In the same model, Tomeglovir at 25 mg/kg b.i.d. for 8 days reduced MCMV DNA in target organs: liver (to 0.2% of placebo), spleen (to 0.6% of placebo), salivary gland, and kidney. Weight loss was reduced to 31.2±10.4% compared to 100±12.5% in placebo. Uninfected mice treated similarly did not develop weight loss. Ganciclovir showed comparable efficacy. [2]

A total of 100 μL of EMEM/10 is added to each well of 96-well microtitre plates to assess the toxicity of the drugs. Following the addition of 2 μL of 50 mM Tomeglovir stock solutions in duplicate into 198 μL in row 2, 100 μL up to row 12 and 100 μL of a HELF, NHDF, or 3T3 cell suspension (5 × 103 cells/mL) are added per well in a series of two-fold dilutions.Row 1 is the control cell that has not been treated. Following a 6-day incubation period at 37°C and 5% CO2, the cells are rinsed once with phosphate-buffered saline (PBS) and 200 μL of fluorescein diacetate, a fluorescent dye solution containing 10 μg/mL in PBS at pH 7.2, are added to each well. A Fluorskan Ascent fluorimeter (emission filter: 530 ± 15 nm, excitation filter: 485 ± 11 nm) is used to measure the fluorescence signal after 45 minutes. CC50 values are calculated graphically, and the relative fluorescence units (RFUs) of treated cells are expressed as percentages of untreated cell controls[2].

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HCMV cytopathogenicity assay (6-day, neutral red staining) or 15-day fluorescence assay: HELF or NHDF cells were seeded in 96-well plates with drug dilutions (250-0.0005 μM), infected with HCMV (moi 0.001-0.002), incubated 6 or 15 days, stained with neutral red or fluorescein diacetate, and EC50/CC50 determined. [2]
Plaque reduction assay: Confluent HELF or NHDF cells in 24-well plates were infected with 40-60 pfu HCMV, overlaid with 0.5% methylcellulose/EMEM/10 containing drug, incubated 7-12 days with medium changes every 3-4 days, stained with neutral red, plaques counted. [2]
For MCMV: similar procedures using MEF or NIH 3T3 cells, moi 0.05-0.1, incubation 5 days, Giemsa staining. [2]
Cytotoxicity fluorescence assay: Cells in 96-well plates with drug dilutions incubated 6 days, washed, then 10 μg/mL fluorescein diacetate added, fluorescence measured (excitation 485 nm, emission 530 nm). CC50 determined graphically. [2]
Southern blot analysis of viral DNA: HELF cells infected with HCMV AD169 (moi 1) with various drug concentrations. Viral DNA isolated, digested with KpnI, size-fractionated, blotted, hybridized with DIG-labeled terminal DNA probe (genome position 703-1524). Chemiluminescence detection. [2]

Mice [1]
Avertin 2.5% (0.015-0.017 mL/g body weight) is used to anesthetize NOD/LtSz-scid/j mice weighing 20–30 g. Avertin 100% is made up of 10 g tribromoethyl alcohol in 10 mL tertiary amyl alcohol. The abdomens are opened and the fibers are inserted intra-abdominally after the belly has been shaved and cleaned aseptically. Two layers of sutures are used to close the abdomens. In this study, only asymptomatic animals are involved. The mice receive Tomeglovir treatment at the recommended dosages twice a day for four days in a row starting the day after transplantation. Under these circumstances, day 5 viral peak titers are seen in preliminary experiments[1].

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MCMV infection model in male NOD/Ltsz-SCID mice (5-6 weeks old). Mice were infected intraperitoneally with 5×10^5 pfu of MCMV strain Smith in 0.2 mL. Approximately 20 h post-infection, Tomeglovir was administered as a suspension in 0.5% Tylose (vehicle) by oral gavage twice daily for 8 consecutive days. Doses: 3, 10, 30, 100 mg/kg body weight per application (total volume 10 mL/kg). Animals were sacrificed 16 h after the last application; organs removed for analysis. [2]
For the 25 mg/kg b.i.d. efficacy study: similar protocol, n=10 per group, treatment started 20 h post-infection, 8 days, then weight loss and MCMV DNA in liver and spleen measured. [2]
Acute toxicity study: oral LD50 in mice and rats >2000 mg/kg. [2]

In mice: oral bioavailability not explicitly stated but the compound is orally active. Plasma protein binding not reported. Clearance not reported. Half-life not reported. However, the compound is well absorbed orally. [2]
In rats and dogs: pharmacokinetic data mentioned but not detailed in this paper; referenced to other abstracts. [2]

Acute oral toxicity: LD50 >2000 mg/kg body weight in mice and rats. [2]
In the efficacy study at 25 mg/kg b.i.d. for 8 days, uninfected mice treated similarly did not develop weight loss. [2]
Cytostatic concentration (CC50) in human NHDF cells: 93 μM; in murine 3T3 cells: 63 μM (as shown in Figure 1). Selectivity indices: HCMV ~300, MCMV ~1600. [2]
No significant weight loss or other overt toxicity reported in treated animals. [2]

[1]. Inhibition of murine cytomegalovirus and human cytomegalovirus by a novel non-nucleosidic compound in vivo. Antiviral Res. 2001 Mar;49(3):179-89.

[2]. Novel non-nucleoside inhibitors of cytomegaloviruses (BAY 38-4766): in vitro and in vivo antiviral activity and mechanism of action. J Antimicrob Chemother. 2001 Dec;48(6):757-67.

Tomeglovir selectively inhibits cytomegaloviruses by targeting the viral DNA cleavage and packaging machinery, specifically the UL89 and UL104 gene products (terminase complex). This mechanism is distinct from DNA polymerase inhibitors (ganciclovir, cidofovir, foscarnet) and no cross-resistance was observed. The compound does not inhibit viral DNA synthesis but blocks the maturation of concatemeric DNA into monomeric genomes, leading to production of empty non-infectious particles. This unique mechanism may allow for potential immune stimulation due to continued viral protein synthesis. [2]
The compound is orally active with excellent tolerability in preclinical species. It is active against HCMV clinical isolates resistant to ganciclovir, foscarnet, and cidofovir. [2]

C23H27N3O4S

441.54318

441.172

C, 62.56; H, 6.16; N, 9.52; O, 14.49; S, 7.26

233254-24-5

475330

Light yellow to yellow solid powder

1.3±0.1 g/cm3

1.667

2.8

3

6

7

31

711

0

CC(C)(CO)C(NC1=CC=C(NS(=O)(C2=C3C=CC=C(N(C)C)C3=CC=C2)=O)C=C1)=O

OSQAKHSYTKBSPB-UHFFFAOYSA-N

InChI=1S/C23H27N3O4S/c1-23(2,15-27)22(28)24-16-11-13-17(14-12-16)25-31(29,30)21-10-6-7-18-19(21)8-5-9-20(18)26(3)4/h5-14,25,27H,15H2,1-4H3,(H,24,28)

N-(4-(((5-(Dimethylamino)-1-naphthyl)sulfonyl)amino)phenyl)-3-hydroxy-2,2-dimethylpropionamide

Tomeglovir; BAY-38-4766; BAY 38-4766; BAY38-4766; BAY-384766; BAY 384766; BAY384766; BAY-38,4766; BAY 38,4766; BAY38,4766.

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

DMSO : ≥ 108 mg/mL (~244.60 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.)