METHODS OF SCREENING PHARMACEUTICALS FOR THE TREATMENT OF PAIN

摘要

Using sphingosine-1-phosphate (S1P), its fragments or derivatives in the preparation of pharmaceutical compounds, for the modulation of pain and for identifying compounds that modulate pain, is new. Independent claims are also included for: (1) screening pharmaceuticals useful for modulating or preventing pain involving: either providing a sample containing protein associated with Myc (PAM), its functional fragment or derivative and not containing S1P, providing a second sample containing PAM, its functional fragment or derivative and containing S1P, measuring the PAM activity in both samples, contacting the first sample with a compound, and determining the ability of the compound to mimic S1P function; or providing two samples comprising a cell expressing an S1P receptor, its functional fragment or derivative, contacting one sample with the compound and measuring or comparing the receptor activity in both samples; or providing two samples comprising a cell expressing S1P receptor, its functional fragment or derivative, contacting the samples with S1P, contacting one sample with the compound and measuring the receptor activity or the interaction of S1P and receptor in both samples; or: (a) providing a cell expressing S1P and PAM or their functional fragment or derivative; (b) contacting the cell with a compound; (c) determining the S1P activity in the presence or absence of compound; and (d) comparing the S1P activity; and (2) identification of a compound that modulates pain involving selecting a compound that modulates or mimics the activity of S1P as a test compound, administering the test compound to a subject, and determination of pain modulation in the subject. ACTIVITY : Analgesic; Antiinflammatory. To determine the analgesic effect of sphingosine-1-phosphate (S1P). S1P was delivered to the spinal cord by intrathecal injection. Rats were anesthetized with ketamine (60 mg/kg intraperitoneally) and midazolam (0.5 - 1 mg/kg intraperitoneally). The skin was incised and catheter was fixed with cyanacrylate glue. Three days after surgery rats were placed into a freely moving system, and 10 MicroM S1P (20 Microl) infused through the catheter. Within 15 minutes after stopping the infusion the formalin test was performed. 5% Formaldehyde solution (50 Microl) was injected subcutaneously. Flinches were counted in one minute intervals up to 60 minutes starting right after formalin injection. Flinches of 5 minutes intervals were counted over a period of 60 minutes. A significant decrease in the number of nociceptive responses for phase 2A (20 - 35 minutes after formalin injection) and detected as compared to phosphate buffer saline/dimethylsulfoxide-treated animals. The results showed that the exogenous S1P acts as an analgesic. No specific results are given. MECHANISM OF ACTION : Adenylyl cyclase (AC) (preferably type I, V or Vf) activity inhibitor; Sphingosine-1-phosphate (S1P) receptor binder.