Physicochemical Properties
| Molecular Formula | C20H25FN2O3S |
| Molecular Weight | 392.49 |
| Exact Mass | 392.157 |
| Elemental Analysis | C, 61.20; H, 6.42; F, 4.84; N, 7.14; O, 12.23; S, 8.17 |
| CAS # | 625820-83-9 |
| PubChem CID | 9952446 |
| Appearance | Typically exists as solid at room temperature |
| LogP | 5.088 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 7 |
| Heavy Atom Count | 27 |
| Complexity | 589 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | S(C(C)C)(NC[C@@](C)(C1C=CC(C2C=CC(C(NC)=O)=CC=2)=CC=1)F)(=O)=O |
| InChi Key | MFJKNXILEXBWNQ-FQEVSTJZSA-N |
| InChi Code | InChI=1S/C20H25FN2O3S/c1-14(2)27(25,26)23-13-20(3,21)18-11-9-16(10-12-18)15-5-7-17(8-6-15)19(24)22-4/h5-12,14,23H,13H2,1-4H3,(H,22,24)/t20-/m0/s1 |
| Chemical Name | 4-[4-[(2R)-2-fluoro-1-(propan-2-ylsulfonylamino)propan-2-yl]phenyl]-N-methylbenzamide |
| Synonyms | LY-503430; LY 503430; 4-{4-[(2R)-2-fluoro-1-(propane-2-sulfonamido)propan-2-yl]phenyl}-N-methylbenzamide; 4-(4-((2R)-2-fluoro-1-(propane-2-sulfonamido)propan-2-yl)phenyl)-N-methylbenzamide; ly 503,430; 625820-83-9; 4-[4-[(2R)-2-fluoro-1-(propan-2-ylsulfonylamino)propan-2-yl]phenyl]-N-methylbenzamide; LY-503,430; LY503430 |
| HS Tariff Code | 2934.99.9001 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets | AMPA receptor PAM/positive allosteric modulator |
| ln Vitro |
Results indicated that submicromolar concentrations of LY503430 selectively enhanced glutamate-induced calcium influx into human embryonic kidney 293 cells transfected with human GLUA1, GLUA2, GLUA3, or GLUA4 AMPA receptors. The molecule also potentiated AMPA-mediated responses in native cortical, hippocampal, and substantia nigra neurons [1].
In Vitro Effects at Cloned Human Inotropic Glutamate Receptors (iGluRs) [2] LY503430 has been tested on human cloned iGluR receptors expressed in HEK293 cells as described by Murray et al. The compound enhanced glutamate-induced calcium influx at recombinant human GLUA1–4 receptors. LY503430 was markedly more potent on “flip” splice variants and showed greater potency on GLUA2 and GLUA4 than on other subunits (Table 1). LY503430 had no effects on kainate-mediated responses in HEK293 cells transfected with GLUK5, GLUK6, or GLUK6K2 subunits. In Vitro Effects on Native Tissue Preparations [2] LY503430 potentiated AMPA-evoked responses in a range of rat neuronal preparations (substantia nigra, cortical, Purkinje, hippocampal, and striatal neurons). When applied in the presence of AMPA, LY503430 produced concentration-dependent potentiation of the evoked current in substantia nigra (Fig. 2A) and striatal giant aspiny neurons (Fig. 2B). The EC50 values for LY503430 were estimated to be 2.7 ìM in substantia nigra and 1.7 ìM in giant aspiny neurons, respectively. Cross Reactivity with Other Neurotransmitter Receptors [2] LY503430 had no effect on sodium, calcium, and potassium channels measured using patch clamp electrophysiology in acutely isolated cortical neurons. In addition, LY503430 was profiled on 18 different neurotransmitter receptors using radiolabeled receptor binding assays. Assays were performed according to standard procedures available in the literature. Membrane homogenates obtained from frozen rat brain tissue or commercially available transfected cell lines were used as a receptor source. Results indicated that at 10 ìM LY503430 had no affinity for the receptors listed in Table 1. Effects on BDNF Levels in Cortical Neurons [2] In addition to their crucial role in synaptic transmission, activation of AMPA receptors has been reported to increase the expression of brain derived neurotrophic factor (BDNF) in vitro and in vivo. This effect appears to be mediated by voltage-gated L-type calcium channels, activated as a consequence of the AMPA receptor-induced membrane depolarization, and by activation of Lyn, a member of the src-family of protein tyrosine kinases, which can physically associate with AMPA receptor subunits. LY503430 produced a large increase in BDNF levels (400 to 1000%) when applied to cortical neurons (Fig. 3), but in most experiments LY503430 had a bell-shaped dose-response curve. Similar increases in BDNF expression were also observed in hippocampal cultures treated with LY503430. |
| ln Vivo |
In the present study, researchers first characterized a novel AMPA receptor potentiator, (R)-4'-[1-fluoro-1-methyl-2-(propane-2-sulfonylamino)-ethyl]-biphenyl-4-carboxylic acid methylamide (LY503430), on recombinant human GLUA1-4 and native preparations in vitro and then evaluated the potential neuroprotective effects of the molecule in rodent models of Parkinson's disease. Researchers also report here that LY503430 provided dose-dependent functional and histological protection in animal models of Parkinson's disease. The neurotoxicity after unilateral infusion of 6-hydroxydopamine into either the substantia nigra or the striatum of rats and that after systemic 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in mice were reduced. Interestingly, LY503430 also had neurotrophic actions on functional and histological outcomes when treatment was delayed until well after (6 or 14 days) the lesion was established. LY503430 also produced some increase in brain-derived neurotrophic factor in the substantia nigra and a dose-dependent increases in growth associated protein-43 (GAP-43) expression in the striatum. Therefore, researchers propose that AMPA receptor potentiators offer the potential of a new disease modifying therapy for Parkinson's disease [1].
In Vivo Effects of LY503430 on Prefrontal Cortical Neurons [2] LY503430 (0.01–10 ìgkg i.v.), potentiated the probability of spike discharge evoked in prefrontal cortical (PFC) neurons by submaximal stimulation of the ventral subiculum in a dose-dependent manner. The threshold dose (0.1 ìgkg) for potentiation by LY503430 of synaptic responses in PFC (Fig. 7) was identical to that for potentiation of iontophoretically applied AMPA-evoked firing of CA1 hippocampal neurons. In contrast to the effects on synaptic responses, LY503430 did not significantly enhance the spontaneous activity of PFC neurons. Furthermore, by systemic administration LY503430 (0.01–10 ìgkg, i.v.) also enhanced responses of hippocampal neurons to iontophoretically applied AMPA in a dose-dependent manner. These data indicate that the brain levels of LY503430 after i.v. administration of the drug at the above listed doses were sufficiently high to produce functional effects. Acute effects on rotational behavior [2] LY503430 (0.5 mgkg s.c.) was evaluated for acute effects on baseline rotational behavior after a unilateral nigral 6-hydroxydopamine (6-OHDA) lesion in rats. In all experiments 4 ìg of 6-OHDA was infused. At this dose 6-OHDA produced a large loss in dopaminergic neurons within 4 to 7 days. In all cases the rats were returned to home cages for 14 days before behavioral testing. Only rats that showed good responses to amphetamine or apomorphine were used in the assessment of new drugs (including LY503430). Acute treatment with LY503430 (0.5 mgkg s.c.) had no effect on asymmetry scores (Fig. 8B). The effects have been compared with those of amphetamine in the same animals (Fig. 8B). In order to assess the effects of LY503430 on rotational behavior during dopaminergic stimulation, the acute effects of the drug in combination with L-DOPA or pergolide were evaluated. Baseline rotations were measured and the rats then received L-DOPA or pergolide and 20 min later were given LY503430 and rotations measured for a further 110 min. The results indicated that both L-DOPA and pergolide produced a robust rotational response, and that this response was not altered by LY503430 (Fig. 9). In a final series of studies unilateral lesioned rats were sensitized with amphetamine and the effects of LY503430 studied after 7 days of sensitization. Data indicated that at 0.5 mgkg LY503430 did not alter amphetamine-sensitized rotational behavior (Fig. 10). Recently, Hess and co-workers reported that AMPA potentiators (CX546 and CX614) from a different class were able to alter methamphetamine-induced circling behavior in methamphetamine-sensitized rats. This is of interest, since it suggests that two different chemical classes of potentiators (from Cortex and Lilly) may have different functional effects in some brain areas. Taken together, the data suggest that LY503430 has no acute effects on rotational behavior. This is not unexpected as there is little evidence that acute activation of AMPA receptors increases dopamine release. Therefore, LY503430 would not be expected to have any acute symptomatic effects in PD. Effects in a mouse MPTP model [2] In 1979, several young adults in the California region were hospitalized with motor problems of an unknown cause: resting tremor, bradykinesia, rigidity, and postural instability. These symptoms are classic hallmarks of Parkinson’s disease and they responded well to L-DOPA, the primary therapy used in PD. It was subsequently discovered that these individuals had received MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) as a contaminant of the drugs they were abusing. It was then determined that MPTP is a potent neurotoxin even at low concentrations. Post-mortem studies carried out when some of those afflicted died from drug overdose revealed that damage to the brain induced by MPTP was remarkably similar to the neurodegeneration observed in PD. Since then many models of PD have been developed using MPTP administered to mice or nonhuman primates (6,46,52). The majority of MPTP animal models use acute administration at doses such as 4 injections of 20 mgkg (in mice) over one day to produce damage to nigrostriatal dopaminergic neurons and achieve a model of end-phase PD within a few days. We used this model to assess the effects of LY503430 and a related compound LY404187. LY503430, daily for 11 days at 0.5 mgkg s.c., prevented the loss of tyrosine hydroxylase immunoreactivity (TH-IR) in the striatum (Fig. 11A) and substantia nigra (Fig. 11B) of MPTP-treated mice. Since it has been reported that the severe dopamine depletion in this “acute” MPTP model is transient and animals recover over time, we also developed a “subchronic” model of MPTP in which the toxin is administered at 30 mgkg for 8 days. Using this model we found a significant protective action of LY503430 that was administered using the same protocol. These data provide evidence that LY503430 can be neuroprotective in various mouse MPTP-induced neurotoxicity models. Effects in a retrograde 6-OHDA model [2] LY503430 was evaluated for its ability to protect against damage to nigral cells following infusion of 6-OHDA into the striatum. In the initial studies we found that 10 ìg of 6-OHDA infused unilaterally into the striatum produces a slow, partial retrograde degeneration of the cell bodies in the substantia nigra resulting in an approximate 50% loss in tyrosine hydroxylase (TH) positive cells at 4 weeks and marked ipsiversive rotations in response to amphetamine (5 mgkg i.p.). This partial retrograde model is thought to mimic some aspects of PD as it has a slow progressive nature. Using this model, we found that dosing with LY503430 (0.5 mgkg s.c.) for 28 days attenuated amphetamine-induced ipsiversive rotations and provided significant protection against the loss of TH positive nigral cell bodies (Fig. 12). Similar protection was observed with the related AMPA receptor potentiator LY404187 suggesting that this class of potentiators is capable of providing neuroprotection in this model. Effects in unilateral 6-OHDA nigral lesion model [2] Our laboratory had also established a more severe 6-hydroxydopamine model, and LY503430 was evaluated in several studies using this model In initial studies, 4 ìg of 6-OHDA infused into the substantia nigra produced a loss of cell bodies over the next 4 days and striatal terminals over the next 5 to 6 days, resulting in an 85–90% loss in nigra cell bodies, 80 to 90% loss of TH-IR in the dorsal striatum and 50–60% loss of TH-IR in the ventral striatum. Subcutaneous and oral efficacy studies [2] Using this model, we initially demonstrated that LY503430 (or LY404187) administered at 0.5 mgkg s.c. for 14 days starting one day after 6-OHDA was able to provide functional improvements (Fig. 13A) and associated preservation of striatal dopaminergic terminals (Fig. 13B). We then carried out a series of experiments to evaluate the effects of LY503430 (0.05, 0.1, 0.2, and 0.5 mgkg p.o. for 10 days, starting on the first day after 6-OHDA) on functional outcome at 12 to 14 days and histological outcome at 13 to 15 days after 6-OHDA. Results of the first experiment indicated that at 0.2 or 0.5 mgkg p.o. LY503430 prevented apomorphine-induced rotations (Fig. 14A) and provided significant protection in the dorsal and ventral striatum (Fig. 14B). Further examination of the doseresponse studies indicated that the minimal effective dose was 0.08 mgkg p.o. These effects were accompanied by only a modest effect on the number of TH positive cells in the substantia nigra. Effects of delayed treatments with LY503430 [2] The large functional improvements and preservation of striatal terminals suggested that the compound had a trophic mechanism of action. To explore this further we carried out an additional series of experiments in which the treatment with LY503430 was delayed for various time intervals after 6-OHDA. Results indicated that LY503430 was able to provide functional and histological benefits when administration began at 3 or 6 days after infusion of 6-OHDA (Fig. 15). There was also a significant effect when treatment was started at 14 days after lesion. These data suggest that LY503430 is acting on either the remaining substantia nigra neurons, the striatum or other brain regions, to activate these areas or to promote reinnervation of the terminal dopaminergic areas. Effects on growth factors and neurite outgrowth [2] Several adjacent sections of striatum were harvested from the oral dose-response and efficacy studies mentioned above. These sections were immunostained for a range of growth factors and growth associated protein–43 (GAP-43), a marker of neurite outgrowth. Results indicated that there were no significant differences in growth factor expression in the striatum between 6-OHDA lesioned animals treated with vehicle or with LY503430 at 12 days. However, there was a dose-dependent increase in GAP-43 in the lesioned striatum with LY503430 (Fig. 16). This finding suggests that new terminals have been formed and explains the large improvement in functional outcome. We also observed some increases in BNDF expression in the substantia nigra after LY503430 and in the hippocampus after LY404187. It is possible that to further understand the mechanisms involved more detailed evaluation of transcription and growth factors at earlier time points is required. Washout study with LY503430 [2] In another experiment we administered LY503430 for 14 days starting one day after 6-OHDA and then stopped treatment (washout). Animals returned to their home cages and were left untreated for the following 28 days prior to evaluating the functional and histological outcome. Using this experimental design we observed significant functional and histological improvements (Fig. 17), suggesting that chronic dosing with LY503430 increases terminal outgrowth and that this effect persists for some time. The data do not mean, however, that only a short period of treatment is required for long lasting effects in human PD, because in the current study the toxin was administered once and then removed from the system, while in PD there is a progressive degeneration. |
| Cell Assay |
Potency and Selectivity of LY503430 [1] Effects on Recombinant GLUA1–4 Receptors Expressed in HEK293 Cells. The AMPA potentiator used for this investigation was the biarylpropylsulfonamide LY503430 (Fig. 1). Submicromolar concentrations of LY503430 enhanced glutamate-induced calcium influx into HEK293 cells transfected with human GLUA1, GLUA2, GLUA3, or GLUA4 AMPA receptors. The potency and efficacy of potentiation by LY503430 were highly dependent on receptor subtype and splice variant. The rank order of potency of LY503430 was GLUA2... |
| ADME/Pharmacokinetics |
Rats [2] The oral bioavailability of LY503430 in rats is 84% (Table 2, Fig. 4). Male F344 rats (n = 3) were dosed orally with 3 mgkg of LY503430 in a 0.5% sodium carboxymethyl cellulose0.25% Tween 80water vehicle, and intravenously with 0.5 mgkg in a 5% Solutol5% ethanol5% propylene glycol85% water vehicle. Plasma was collected at regular intervals over 24 h after treatment. The plasma elimination half-life was 1.2 hours after intravenous and 3 to 4 h after oral treatment. Rat brain levels of LY503430 were approximately 20% of the plasma levels over the eight hour post-dosing interval as shown in Table 3 and Fig. 5. The half-life of LY503430 in brain was 1 h. Dogs [2] The oral bioavailability of LY503430 in dogs is 66 ± 15% (Table 4, Fig. 6). Dogs (2 males and 1 female) were dosed orally with 1 mgkg of LY503430 in a 0.5% sodium carboxymethyl cellulose0.25% Tween 80water vehicle, and intravenously with 0.1 mgkg of LY503430 in a 5% Solutol5% ethanol5% propylene glycol85% water vehicle. Plasma was collected over the 0.5–24 h post dosing interval. The mean plasma elimination half-life was two hours after intravenous dosing and 4.7 h after oral dosing |
| References |
[1]. LY503430, a novel alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor potentiator with functional, neuroprotective and neurotrophic effects in rodent models of Parkinson's disease. J Pharmacol Exp Ther. 2003 Aug;306(2):752-62. [2]. LY503430: Pharmacology, Pharmacokinetics, and Effects in Rodent Models of Parkinson's Disease. CNS Drug Rev. 2006 Jun 7;11(1):77–96. |
| Additional Infomation |
Glutamate is the major excitatory transmitter in the brain. Recent developments in the molecular biology and pharmacology of the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) subtype of glutamate receptors have led to the discovery of selective, potent, and systemically active AMPA receptor potentiators. These molecules enhance synaptic transmission and play important roles in plasticity and cognitive processes. [1]
In the present study, we have demonstrated that a novel, potent, selective, and systemically active AMPA receptor potentiator provided neuroprotective actions in three rodent models of Parkinson’s disease. The compound also increased GAP-43 expression in the lesioned striatum and reduced the 6-OHDA-induced effects on turning behavior and on striatal DAergic innervation. LY503430 is a new class of compound with potential utility for providing neuroprotection and neuronal repair in Parkinson’s...[1] Glutamate is the major excitatory transmitter in the brain. Recent developments in the molecular biology and pharmacology of the α‐amino‐3‐hydroxy‐5‐methylisoxa‐zole‐4‐propionic acid (AMPA)‐subtype of glutamate receptors have led to the discovery of selective, potent and systemically active AMPA receptor potentiators. These molecules enhance synaptic transmission and play important roles in plasticity and cognitive processes. In the present studies we characterized a novel AMPA receptor potentiator, LY503430, on recombinant human GLUA1‐4 and native preparations in vitro, and then evaluated the potential neuroprotective effects of the molecule in rodent models of Parkinson's disease. Results indicated that at submicromolar concentrations LY503430 selectively enhanced glutamate‐induced calcium influx into HEK293 cells transfected with human GLUA1, GLUA2, GLUA3, or GLUA4 AMPA receptors. The molecule also potentiated AMPA‐mediated responses in native cortical, hippocampal and substantia nigra neurones. LY503430 had good oral bioavailability in both rats and dogs. We also report here that LY503430 provided dose‐dependent functional and histological protection in animal models of Parkinson's disease. The neurotoxicity following unilateral infusion of 6‐hyrdoxydopamine (6‐OHDA) into either the substantia nigra or the striatum of rats and that following systemic 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP) in mice were reduced. Interestingly, LY503430 also had neurotrophic actions on functional and histological outcomes when treatment was delayed until well after (6 or 14 days) the lesion was established. LY503430 also produced some increase in brain derived neurotrophic factor (BDNF) in the substantia nigra and a dose‐dependent increase in growth associated protein‐43 (GAP‐43) expression in the striatum. Therefore, we propose that AMPA receptor potentiators such as LY503430 offer the potential of a new disease modifying therapy for Parkinson's disease. [2] Parkinson’s disease (PD) is a progressive, neurodegenerative disorder of the basal ganglia. There is a large loss of dopaminergic cells in the substantia nigra, which project to the terminal rich caudate and putamen of the corpus striatum. Clinical symptoms are manifested when approximately 60% of cell bodies have degenerated and initial symptoms of PD include tremor at rest, muscular rigidity, bradykinesia, postural abnormalities and instability. The available pharmacotherapies involve dopamine replacement. These drugs reduce symptom severity, but do not dramatically affect disease progression. Therefore, in order to maintain an acceptable quality of life for patients with PD, therapies that slow or stop disease progression are needed. A variety of growth factors, including glial derived neurotrophic factor (GDNF) and BDNF, have been reported to promote the survival of dopaminergic neurons in culture and to protect against neurotoxin-induced lesions of the nigrostriatal system. A major drawback of growth factors as therapeutic agents is the need for their central application. Many investigators have reported that antioxidants, nitric oxide synthase inhibitors, antiinflammatory agents, nicotine, immunophilins, and related molecules can provide protection in rodent models of PD. Another approach would be to boost endogenous neurogenesis, plasticity and growth factor expression with small molecules. The data presented in this review suggest that AMPA receptor potentiators such as LY503430 may act by this mechanism. It is possible that the reinnervation and sprouting of the dopamine-depleted striatum in the rodent model may also occur in PD patients. If so, an agent such as LY503430 while not producing any acute symptomatic effect may over time increase striatal innervation, slow disease progression and improve function. Other recent studies have reported that AMPA can protect cultured neurons against glutamate excitotoxicity through a phosphatidylinositol 3-kinase-dependent activation of extracellular signal-regulated kinase leading to upregulation of BDNF gene expression and that AMPA receptor potentiators are neuroprotective against lesions in neonatal mouse brain. Advances in imaging techniques have allowed clinical trials to assess disease progression. The REAL PET study suggested that there was slower progression of Parkinson’s disease with ropinirole versus levodopa. The technology can also be used to visualize new dopaminergic terminals following infusion of GDNF in Parkinson’s disease and functional receptors after fetal dopaminergic transplantation. It should, therefore, be possible to study the disease modifying (neuroprotective) or enhanced sprouting (neurotrophic) actions with agents such as LY503430 in the clinical situation. [2] |
Solubility Data
| Solubility (In Vitro) | May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples |
| Solubility (In Vivo) |
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. Injection Formulations (e.g. IP/IV/IM/SC) 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). 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) Oral Formulations 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). 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 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5478 mL | 12.7392 mL | 25.4784 mL | |
| 5 mM | 0.5096 mL | 2.5478 mL | 5.0957 mL | |
| 10 mM | 0.2548 mL | 1.2739 mL | 2.5478 mL |