Biomarkers for sensitive detection of statin-induced muscle toxicity

Abstract

The present invention inter alia provides a method, and uses thereof, of predicting statin-induced muscle toxicity or its complications, such as myalgia, myopathy and rhabdomyolysis, by detecting the lipid concentrations or lipid-lipid concentration ratios of a biological sample and comparing them to a control. This method has identified lipid markers that are more specific and sensitive in detecting these statin-induced muscle toxicity than the currently utilized clinical markers. Also provided is an antibody towards said lipids, and the use thereof for predicting, diagnosing, statin-induced muscle toxicity. The invention additionally relates to kits comprising lipids and/or an antibody thereto, for use in the prediction and/or diagnosis of statin-induced muscle toxicity.

1 .- 31 . (canceled) 
     
     
         32 . A method for determining whether a subject is at risk to develop, or is suffering from muscle toxicity induced by a lipid lowering drug and/or one or more of its complications, comprising
 (a) determining in a sample from said subject the concentration(s) of one or more lipid(s), wherein (an) increased or decreased concentration(s) in said sample, when compared to a control, is (are) indicative of said subject suffering from said muscle toxicity induced by the lipid lowering drug and/or said complication(s), wherein the one or more lipid(s) whose increase in concentration is (are) compared to the control is (are) selected from: LacCer(d18:1/22:0), Gb3(d18:1/24:1), Gb3(d18:1/22:0), Gb3(d18:1/24:0), LacCer(d18:1/24:0), Total eicosanoids, Total Gb3, Cer(d18:1/20:0), Gb3(d18:1/20:0), LacCer(d18:1/24:1), LacCer(d18:1/20:0), PGE2, 12-OXOETE, 17-HDoHE, PGD2, TXB3, PS 18:0/18:1 and SM (d18:1/24:2);   and wherein the one or more lipid(s) whose decrease in concentration is (are) compared to the control is (are) selected from: Gb3(d18:1/16:0) and Gb3(d18:1/18:0); or   (b) determining in a sample from said subject one or more lipid-lipid concentration ratio(s), wherein (an) increased or decreased lipid-lipid concentration ratio(s) in said sample, when compared to a control, is (are) indicative of said subject suffering from said muscle toxicity induced by the lipid lowering drug and/or said complication(s), wherein the one or more lipid-lipid concentration ratio(s) whose increase is (are) compared to the control is (are) selected from: 12-HETE/15-HETrE, 12-HETE/14_15-DHET, 12-HETE/Gb3(d18:1/16:0), 12-HETE/Glc/GalCer(d18:1/24:1), 12-HETE/Glc/GalCer(d18:1/18:0), Gb3(d18:1/24:1)/LacCer(d18:1/16:0), LacCer(d18:1/22:0)/LacCer(d18:1/24:1), Cer(d18:1/20:0)/Glc/GalCer(d18:1/24:1); and wherein the one or more lipid-lipid concentration ratio(s) whose decrease is (are) compared to the control is (are) selected from: LacCer(d18:1/16:0)/LacCer(d18:1/22:0), Gb3(d18:1/16:0)/Gb3(d18:1/24:1) and 11-HETE/12-HETE, and   (c) adjusting a treatment of the subject who is determined to be at risk to develop, or is suffering from muscle toxicity induced by a lipid lowering drug and/or one or more of its complications, wherein adjusting the treatment comprises (i) reducing a lipid lowering drug dose; (ii) stopping a lipid lowering drug treatment; or (iii) changing to a different lipid lowering drug.   
     
     
         33 . A method for determining whether the treatment with a lipid lowering drug of a subject needs adjustment, comprising
 (a) determining in a sample from said subject the concentration(s) of one or more lipid(s), wherein (an) increased or decreased concentration(s) in said sample, when compared to a control, is (are) indicative of said treatment requiring adjustment, wherein the one or more lipid(s) whose increase in concentration is (are) compared to the control is (are) selected from: LacCer(d18:1/22:0), Gb3(d18:1/24:1), Gb3(d18:1/22:0), Gb3(d18:1/24:0), LacCer(d18:1/24:0), Total eicosanoids, Total Gb3, Cer(d18:1/20:0), Gb3(d18:1/20:0), LacCer(d18:1/24:1), LacCer(d18:1/20:0), PGE2, 12-OXOETE, 17-HDoHE, PGD2, TXB3, PS 18:0/18:1 and SM (d18:1/24:2);   and wherein the one or more lipid(s) whose decrease in concentration is (are) compared to the control is (are) selected from: Gb3(d18:1/16:0) and Gb3(d18:1/18:0); or   (b) determining in a sample from said subject one or more lipid-lipid concentration ratio(s), wherein (an) increased or decreased lipid-lipid concentration ratio(s) in said sample, when compared to a control, is (are) indicative of said treatment requiring adjustment, wherein the one or more lipid-lipid concentration ratio(s) whose increase is (are) compared to the control is (are) selected from: 12-HETE/15-HETrE, 12-HETE/14_15-DHET, 12-HETE/Gb3 (d18:1/16:0), 12-HETE/Glc/GalCer(d18:1/24:1), 12-HETE/G1c/GalCer(d18:1/18:0), Gb3(d18:1/24:1)/LacCer(d18:1/16:0), LacCer(d18:1/22:0)/LacCer(d18:1/24:1), Cer(d18:1/20:0)/Glc/GalCer(d18:1/24:1);   and wherein the one or more lipid-lipid concentration ratio(s) whose decrease is (are) compared to the control is (are) selected from: LacCer(d18:1/16:0)/LacCer(d18:1/22:0), Gb3(d18:1/16:0)/Gb3(d18:1/24:1) and 11-HETE/12-HETE, and   (c) adjusting the treatment with the lipid lowering drug in the subject who is determined to need adjustment, wherein adjusting the treatment comprises (i) reducing a lipid lowering drug dose; (ii) stopping a lipid lowering drug treatment; (iii) re-commencing a lipid lowering drug treatment; or (iv) changing to a different lipid lowering drug.   
     
     
         34 . The method of  claim 32 , wherein the lipid whose increase in concentration is compared to the control is selected from: LacCer(d18:1/22:0) and Gb3(d18:1/24:1). 
     
     
         35 . The method of  claim 33 , wherein the lipid whose increase in concentration is compared to the control is selected from: LacCer(d18:1/22:0) and Gb3(d18:1/24:1). 
     
     
         36 . The method of  claim 33 , wherein the adjustment of said treatment with a lipid lowering drug comprises
 (a) a reduction of a statin dose;   (b) a cessation of a statin treatment;   (c) a re-commencement of a statin treatment;   (d) a change to a different statin drug;   (e) a change to a different lipid lowering drug; or   (f) a cessation of another drug treatment which led to muscle toxicity due to its interaction with one or more statins.   
     
     
         37 . The method of  claim 32 , wherein the method is for evaluating the degree of muscle toxicity induced by a novel statin or a novel lipid lowering medication in a subject undergoing treatment with said statin or lipid lowering medication. 
     
     
         38 . The method of  claim 32 , wherein the method is for
 (a) determining early warning signs of muscle toxicity in said subject and/or   (b) determining whether the symptoms of muscle toxicity found in a subject are due to statin-induced muscle toxicity.   
     
     
         39 . The method of  claim 32 , comprising determining at least 2, at least 3, at least 4, or at least 5, of said lipid concentrations or lipid-lipid concentration ratios. 
     
     
         40 . The method of  claim 33 , comprising determining at least 2, at least 3, at least 4, or at least 5, of said lipid concentrations or lipid-lipid concentration ratios. 
     
     
         41 . The method of  claim 32 , wherein said lipid lowering drug is a statin and wherein
 (a) said subject is being treated with one or more statins;   (b) said subject had undergone statin treatment, but discontinued said treatment due to onset of muscle pain; or   (c) said subject has not yet been treated with statins.   
     
     
         42 . The method of  claim 33 , wherein said lipid lowering drug is a statin and wherein
 (a) said subject is being treated with one or more statins;   (b) said subject had undergone statin treatment, but discontinued said treatment due to onset of muscle pain; or   (c) said subject has not yet been treated with statins.   
     
     
         43 . The method of  claim 32 , wherein the subject is at a high risk for developing statin-induced muscle toxicity and/or one or more of its complications. 
     
     
         44 . The method of  claim 33 , wherein the subject is at a high risk for developing statin-induced muscle toxicity and/or one or more of its complications. 
     
     
         45 . The method  claim 32 , wherein said control to which comparison is made is:
 (a) a control sample from the same subject undergoing statin treatment prior to the onset of muscle toxicity;   (b) a control sample from the same subject prior to statin treatment or during discontinuation of statin treatment;   (c) a control sample from a subject with no signs or history of statin-induced muscle toxicity;   (d) a control sample from a population of subjects with no signs or history of statin-induced muscle toxicity;   (e) a control value established from one or more subject(s) not on statin treatment and with no signs or history of muscle toxicity; or   (f) a control value established from one or more subject(s) on statin treatment and with no signs or history of muscle toxicity.   
     
     
         46 . The method  claim 33 , wherein said control to which comparison is made is:
 (a) a control sample from the same subject undergoing statin treatment prior to the onset of muscle toxicity;   (b) a control sample from the same subject prior to statin treatment or during discontinuation of statin treatment;   (c) a control sample from a subject with no signs or history of statin-induced muscle toxicity;   (d) a control sample from a population of subjects with no signs or history of statin-induced muscle toxicity;   (e) a control value established from one or more subject(s) not on statin treatment and with no signs or history of muscle toxicity; or   (f) a control value established from one or more subject(s) on statin treatment and with no signs or history of muscle toxicity.   
     
     
         47 . The method of  claim 32 , further comprising determining or evaluating the level of creatine kinase in said subject or in a sample from said subject. 
     
     
         48 . The method of  claim 47 , wherein the subject has or does not have elevated creatine kinase levels. 
     
     
         49 . The method of  claim 33 , further comprising determining or evaluating the level of creatine kinase in said subject or in a sample from said subject. 
     
     
         50 . The method of  claim 49 , wherein the subject has or does not have elevated creatine kinase levels. 
     
     
         51 . The method of  claim 32 , wherein
 (a) the sample is blood, plasma, serum, a lipoprotein fraction of blood, urine or muscle biopsy tissue; and/or   (b) the lipid concentration(s) or lipid-lipid concentration ratio(s) is (are) determined by using mass spectrometry, nuclear magnetic resonance spectroscopy, fluorescence spectroscopy or dual polarisation interferometry, a high performance separation method such as HPLC or UPLC, an immunoassay such as an ELISA and/or with a binding moiety capable of specifically binding the analyte.   
     
     
         52 . The method of  claim 33 , wherein
 (a) the sample is blood, plasma, serum, a lipoprotein fraction of blood, urine or muscle biopsy tissue; and/or   (b) the lipid concentration(s) or lipid-lipid concentration ratio(s) is (are) determined by using mass spectrometry, nuclear magnetic resonance spectroscopy, fluorescence spectroscopy or dual polarisation interferometry, a high performance separation method such as HPLC or UPLC, an immunoassay such as an ELISA and/or with a binding moiety capable of specifically binding the analyte.   
     
     
         53 . A kit for predicting statin-induced muscle toxicity and/or one or more of its complications, wherein the kit comprises:
 (a) lipid standard(s) chosen from the lipids defined in claim  1 , and optionally one or more further reference compounds selected from:   (b) one or more control markers;   (c) positive and/or negative controls;   (d) internal and/or external standards;   (e) calibration line controls; and   (f) an agent, optionally an antibody, capable of binding any one of the lipids defined in claim  1 .   
     
     
         54 . Use of a kit as defined in  claim 53  for predicting statin-induced muscle toxicity and/or one or more of its complications, wherein the lipid concentration(s) or lipid-lipid concentration ratio(s) in a sample from a subject is (are) optionally determined by using mass spectrometry. 
     
     
         55 . The method of  claim 32 , wherein:
 (a) said statin is selected from the group consisting of atorvastatin, cerivastatin, fluvastatin, fluvastatin XL, lovastatin, pitavastatin, pravastatin, rosuvastatin and/or simvastatin;   (b) said lipid lowering drug may be selected from any of the following: a HMG-CoA reductase inhibitor, niacin (nicotinic acid), a cholesterol absorption inhibitor, a cholesteryl ester transfer protein (CETP) inhibitor, a bile acid sequestrant; a fibrate or a phytosterol;   wherein optionally said cholesterol absorption inhibitor is ezetimibe or SCH-48461; said cholesteryl ester transfer protein (CETP) inhibitor is torcetrapib, anacetrapib or dalcetrapib; said bile acid sequestrant is colesevelam, cholestyramine or colestipol; and said fibrate is fenofibrate, gemfibrozil, clofibrate, or bezafibrate;   (c) the muscle toxicity is associated with a muscle disease, for example, a muscle dystrophy; and/or   (d) the one or more complications of statin-induced muscle toxicity are selected from myalgia, myositis, myopathy and rhabdomyolysis.   
     
     
         56 . The method of  claim 33 , wherein:
 (a) said statin is selected from the group consisting of atorvastatin, cerivastatin, fluvastatin, fluvastatin XL, lovastatin, pitavastatin, pravastatin, rosuvastatin and/or simvastatin;   (b) said lipid lowering drug may be selected from any of the following: a HMG-CoA reductase inhibitor, niacin (nicotinic acid), a cholesterol absorption inhibitor, a cholesteryl ester transfer protein (CETP) inhibitor, a bile acid sequestrant; a fibrate or a phytosterol;   wherein optionally said cholesterol absorption inhibitor is ezetimibe or SCH-48461; said cholesteryl ester transfer protein (CETP) inhibitor is torcetrapib, anacetrapib or dalcetrapib; said bile acid sequestrant is colesevelam, cholestyramine or colestipol; and said fibrate is fenofibrate, gemfibrozil, clofibrate, or bezafibrate;   (c) the muscle toxicity is associated with a muscle disease, for example, a muscle dystrophy; and/or   (d) the one or more complications of statin-induced muscle toxicity are selected from myalgia, myositis, myopathy and rhabdomyolysis.   
     
     
         57 . The method of  claim 32 , wherein the lipid lowering drug is a statin. 
     
     
         58 . The method of  claim 33 , wherein the lipid lowering drug is a statin.