Synthesis, compositions and methods for the measurement of the concentration of stable-isotope labeled compounds in life forms and life form excretory products

Abstract

Stable isotope labeling and neutron activation to measure biological functions are provided, as are the use and method of adding a chemical monitor to correct for neutron flux to sample vials prior to the addition of sample is presented, and the use of stable isotopes as a chemical bar code for vials and other items. Methods are provided also for measuring glomerular filtration rate and glomerular sieving function in a subject, and for measuring other physiological functions.

1 . A composition comprising a compound having a structure X-M, wherein X is a chelator, M is an atom of a stable isotope of an element having a nucleus capable of capturing a neutron and subsequently emitting a photon, M is noncovalently bound to X, X being at a concentration at least as great as that of M, the composition having a counterion and a physiologically acceptable buffer.  
   
   
       2 . A composition according to  claim 1 , wherein M is selected from the group consisting of:  45 Sc,  50 Cr,  55 Mn,  58 Fe,  59 Co,  63 Cu,  103 Rh,  13 Cd,  114 Cd,  113 In,  115 In,  123 Te,  133 Cs,  139 La,  141 Pr,  146 Nd,  149 Sm,  152 Sm,  15 Eu,  153 Eu,  152 Gd,  155 Gd,  157 Gd,  159 Tb,  158 Dy,  160 Dy,  161 Dy,  162 Dy,  163 Dy,  164 Dy,  168 Yb,  169 Tm,  174 Hf,  178 Hf,  175 Yb,  165 Ho,  175 Lu,  176 Lu,  181 Ta,  185 Re, Re,  190 Ir,  193 Ir,  196 Hg,  202 Hg, and  197 Au.  
   
   
       3 . A composition according to  claim 1 , wherein the counterion is meglumine (1-deoxy-1-methylamino-D-glucitol antimoniate).  
   
   
       4 . A composition according to  claim 1 , wherein X is selected from the group consisting of: diethylenetriaminepentaacetic acid (DTPA), diethylenetriamine-pentamethylenephosphonic acid (DTPMP), tetraazacyclododecanetetraacetic acid (DOTA) or a derivative of DOTA, ethylene-diaminetetraacetic acid (EDTA), tetraazacyclododecanetetrakis (methylene phosphonic acid) (DOTP), hydroxypropyl tetraazacylododecanetriacetic acid (HP-DOPA), diethylenetriaminetriacetic acid bismethylamide (DTPA-BMA), and MS-325.  
   
   
       5 . A composition according to  claim 1 , wherein the concentration of X is between about 100 micromolar and about 1.5 molar.  
   
   
       6 . A method of preparing a pharmaceutical composition of a compound having a structure X-M, the method comprising: 
 providing a solution having X and M, wherein X is a chelator, and M is an atom of a stable isotope of an element noncovalently bound to X and having a nucleus capable of capturing a neutron and subsequently emitting a photon, and the concentration of X is at least as great as that of M, the solution being aqueous and having a physiologically acceptable buffer; and    sterilizing the solution.    
   
   
       7 . A method according to  claim 6 , wherein M is selected from the group consisting Of  45 Sc  50 Cr, 55Mn,  58 Fe,  59 Co,  63 Cu,  103 Rh,  113 Cd,  114 Cd,  113 In,  115 In,  123 Te,  133 Cs,  139 La,  141 Pr,  146 Nd,  149 Sm,  152 Sm,  151 Eu,  153 Eu,  152 Gd,  155 Gd,  157 Gd,  159 Tb,  158 Dy,  160 Dy,  161 Dy,  162 Dy,  163 Dy,  164 Dy,  168 Yb,  169 Tm,  174 Hf,  178 Hf,  175 Yb,  165 Ho,  175 Lu,  176 Lu,  181 Ta,  185 Re,  187 Re,  190 Ir,  193 Ir,  196 Hg,  202 Hg, and  197 Au.  
   
   
       8 . A method for quantifying by neutron activation an amount of a complex having a structure X-M, the method comprising: 
 exposing each of the X-M complex and a standard to a neutron source, wherein X is a chelator and M is an atom of a stable isotope of an element having a nucleus capable of capturing a neutron and subsequently emitting a photon, and wherein the standard has a predetermined quantity of M, the X-M and the standard being exposed to the neutron source at the same time, such that M emits a photon after capture of a neutron;    detecting an emitted photon from each of X-M and the standard; and    comparing an amount of photonic emissions from each of the X-M and the standard, thereby quantifying the amount of the complex having the structure X-M.    
   
   
       9 - 22 . (canceled)  
   
   
       23 . A composition comprising at least one compound Ω i -X j -M k  and a counterion, wherein: each Ω i  is an organic compound having a molecular weight greater than about 50; each X j  is at least one chelator covalently bound to each Ω i ; and each M k  is noncovalently bound to X and has a nucleus capable of capturing a neutron and subsequently emitting a photon and is an atom of a stable isotope of an element selected from the group consisting of  45 Sc,  50 Cr,  55 Mn,  58 Fe,  59 Co,  63 Cu,  103 Rh,  113 Cd,  114 Cd,  113 In,  115 In,  123  Te,  133 Cs,  139 La,  141 Pr,  146 Nd,  149 Sm,  152 Sm,  151 Eu,  153 Eu,  152 Gd,  155 Gd,  157 Gd,  159 Tb,  158 Dy,  160 Dy,  16 Dy,  162 Dy,  163 Dy,  164 Dy,  168 Yb,  169 Tm,  174 Hf  178 Hf,  175 Yb,  165 Ho,  175 Lu,  176 Lu,  181 Ta,  185 Re,  187 Re,  190 Ir,  193 Ir,  196 Hg,  202 Hg, and  197 Au, wherein each M k  is distinct in identity; i, j, and k each being a number from 1 to 8, wherein i is at least equal to k.  
   
   
       24 . A composition comprising at least one compound Ω i -X j -Mk, at least one compound Ψ-N, and a cationic counterion, wherein: each Ω is an organic compound having a molecular weight greater than about 50; each X is at least one chelator covalently bound to Ω and noncovalently bound to M; Ψ is a chelator noncovalently bound to N; N is different from M, and N and each M being, independently, an atom of a stable isotope of an element selected from the group consisting of  45 Sc,  50 Cr,  55 Mn,  58 Fe,  59 Co,  63 Cu,  103 Rh,  113 Cd,  114 Cd,  113 In,  115 In,  123 Te,  133 Cs,  139 La,  141 Pr,  146 Nd,  149 Sm,  152 Sm,  151 Eu,  153 Eu,  152 Gd,  155 Gd,  157 Gd,  159 Tb,  158 Dy,  160 Dy,  161 Dy,  162 Dy,  163 Dy,  164 Dy,  168 Yb,  169 Tm,  174 Hf,  178 Hf  175 Yb,  165 Ho,  175 Lu,  176 Lu,  181 Ta,  185 Re,  187 Re,  190 Ir,  193 Ir,  196 Hg,  202 Hg, and  197 Au, each M and N, independently, having a nucleus capable of capturing a neutron and subsequently emitting a photon; and i, j and k are independently each a number from 1 to about 8.  
   
   
       25 . A composition comprising: a compound Ω i -M and a cationic counterion, wherein: each Ω is an organic compound having a molecular weight greater than about 50, i being a number from 1 to about 8; and M is an atom of a stable isotope of an element selected from the group of elements consisting of  36 S,  74 Se,  79 Br,  81 Br,  107 Ag,  109 Ag,  127 I,  197 Au,  190 Pt, and  196 Hg, wherein M is covalently bound to Q and has a nucleus capable of capturing of a neutron and subsequently emitting a photon.  
   
   
       26 . A composition according to  claim 24 , wherein the at least one Ω i -X-M i  compound differs in molecular weight range from the other compounds, the Ω 1 -X-M i  compounds having the same net charge.  
   
   
       27 . A composition according to  claim 24 , wherein the Ω i -X-M i  compounds have different net charges and about the same molecular weight.  
   
   
       28 . A composition according to  claim 24 , wherein Ω i  is a polymer.  
   
   
       29 . A composition according to  claim 28 , wherein the polymer is selected from the group consisting of a polysaccharide, a polypeptide, and a polynucleotide.  
   
   
       30 . A composition according to  claim 29 , wherein the polysaccharide is a ficoll, a dextran, a pullulan, a starch, or a hydroxyethylstarch.  
   
   
       31 . A composition according to  claim 29 , wherein the polypeptide is covalently attached to a polyethylene glycol polymer.  
   
   
       32 . A composition according to  claim 24 , wherein the non-covalently bound chelator is a bile acid compound.  
   
   
       33 . A composition according to  claim 32 , wherein the bile acid compound is selected from the group consisting of cholic acid, cholic acid taurine, chenodeoxycholic, deoxycholic acid, homocholic acid taurine, and lithocholic acid.  
   
   
       34 . A composition according to  claim 32 , wherein the bile acid compound is a synthetic derivative of a bile acid.  
   
   
       35 . A composition according to  claim 24 , wherein the non-covalently bound chelator is a drug or a drug metabolite.  
   
   
       36 . A composition according to  claim 35 , wherein Ω i  is selected from the group consisting of a hormone and a hormone antagonist.  
   
   
       37 . A composition according to  claim 36 , wherein Ω i  is a steroid hormone.  
   
   
       38 . A composition according to  claim 24 , wherein the non-covalently bound chelator is selected from the group consisting of an antibiotic, a tranquilizer, a vitamin, a narcotic, a cannabinoid, a barbiturate, and an alkaloid.  
   
   
       39 . A composition comprising a plurality of colloids having the structure Y—O u -M t  and a cation counterion, the composition being suspended in a physiologically compatible buffer, wherein: Y is a polymer having a molecular weight greater than about 1000; O is oxygen and u is a number between zero and about 200; and each M is an atom selected from the group of stable isotopes capable of capturing a neutron, thereby becoming unstable and emitting a photon having a characteristic energy spectrum, the photon being selected from the group consisting of a gamma photon, an x-ray photon or a prompt photon, t being an integer from 1 to about 10.  
   
   
       40 . A composition according to  claim 39 , wherein each colloid among the plurality has a distinct molecular weight and is uniquely associated with a distinct M.  
   
   
       41 . A method for quantifying by neutron activation an amount of at least one compound Ω i -X j -M k , wherein each Ω i  is a unique organic compound having a molecular weight greater than about 50; each X j  is at least one chelator covalently bound to Ω i ; and each M k  is an atom of a stable isotope of an element and is distinct in identity and has a nucleus capable of capturing neutrons and thereby emitting photons, and i, j, and k are each numbers from 1 to 8, i being at least as great as k, the method comprising: 
 exposing a first container having Ω i -X j -M k  and a second container having a standard known quantity of M k  to a neutron source using the same neutron field, such that Ω i -X j -M k  and the M k  standard capture neutrons and emit photons, wherein each M k  is selected from the group consisting of  45 Sc,  50 Cr,  55 Mn,  58 Fe,  59 Co,  63 Cu,  103 Rh,  113 Cd,  114 Cd,  113 In,  115 In,  123 Te,  133 Cs,  139 La,  141 Pr,  141 Nd,  149 Sm,  152 Sm,  153 Eu,  153 Eu,  152 Gd,  155 Gd,  157 Gd,  159 Tb,  158 Dy,  160 Dy,  161 Dy,  162 Dy,  163 Dy,  164 Dy,  168 Yb,  169 Tm,  174 Hf,  178 Hf,  175 Yb,  165 Ho,  175 Lu,  176 Lu,  181 Ta,  185 Re,  187 Re,  190 Ir,  193 Ir,  196 Hg,  202 Hg, and  197 Au;    detecting resulting photon emissions; and    comparing photon emissions of Ω i -X j -M k  with photon emissions of the standard, thereby quantifying the amount of at least one compound Ω i -X j -M k .    
   
   
       42 . A method of evaluating the rate of at least one physiological process in a subject, the method comprising: 
 administering at least one test composition to a subject, wherein each test composition is differently labeled with one of the group of stable isotopes of one or more elements;    obtaining a plurality of samples of a bodily fluid from the subject, the samples being obtained at different times after administering the composition;    determining by neutron activation the amount of the at least one test composition in a volume of the samples; and    calculating the rate of change of concentration over time of the composition in the bodily fluid, thereby evaluating the rate of the physiological process in the subject.    
   
   
       43 . A method according to  claim 42 , wherein the rate of the physiological process is the glomerular filtration rate.  
   
   
       44 . A method according to  claim 42 , wherein the physiological process is the glomerular integrity rate.  
   
   
       45 . A method according to  claim 42 , wherein the rate of the at least one physiological process is a glomerular filtration rate and a glomerular integrity rate.  
   
   
       46 . A method according to  claim 42 , wherein the rate of change of concentration of the at least one test composition is glomerular selectivity according to size of the test composition.  
   
   
       47 . A method according to  claim 42 , wherein the rate of change of concentration of the at least one test composition is glomerular selectivity according to charge of the test composition.  
   
   
       48 . A method according to  claim 42 , wherein the physiological process is at least one hepatic function.  
   
   
       49 . A method according to  claim 42 , wherein the physiological process is at least one gastrointestinal function.  
   
   
       50 . A method according to  claim 42 , wherein evaluating the rate of the physiological process is evaluating absorption of a bile acid compound.  
   
   
       51 . A method according to  claim 42 , wherein evaluating the rate of the physiological process is evaluating cirrhosis.  
   
   
       52 . A method according to  claim 42 , wherein evaluating the rate of change of the physiological process is evaluating liver function after liver transplantation.  
   
   
       53 . A kit for measuring glomerular integrity rate, comprising: a test composition X-M and a counterion in a physiologically compatible solution wherein X is a chelator noncovalently bound to M, and M is an atom of a stable isotope of an element having a nucleus capable of capturing a neutron and subsequently emitting a photon; at least one sample container to collect at least one samples of a bodily fluid; instructions for use; and a data recording system.  
   
   
       54 . A composition for lot labeling a plurality of sample containers, the composition comprising at least one stable isotope of an element and a binding agent, wherein the binding agent maintains the stable isotope in the container, the stable isotope having a nucleus capable of capturing a neutron and subsequently emitting a photon.  
   
   
       55 . A composition according to  claim 54 , wherein the binding agent is selected from the group consisting of: a polyurethane, a wax, an oil based adhesive, an organic solvent, a water based adhesive, an aqueous solvent, an acrylic and a plastic.  
   
   
       56 . A method for monitoring neutron flux variation within a set of samples, the method comprising: 
 providing a lot having a plurality of sample containers, each container having a predetermined amount of a first stable isotope;    adding a test sample containing a second stable isotope to each of the containers, wherein the first and second stable isotopes are different and are selected from the group consisting Of  45 Sc,  50 Cr,  55 Mn,  58 Fe,  59 Co,  63 Cu,  103 Rh,  113 Cd,  114 Cd,  113 In,  115 In,  123 Te,  133 Cs,  139 La,  141 Pr,  146 Nd,  149 Sm,  152 Sm,  151 Eu,  153 Eu,  152 Gd,  155 Gd,  157 Gd,  159 In,  158 Dy,  160 Dy,  16 Dy,  162 Dy,  163 Dy,  164 Dy,  168 Yb,  169 Tm,  174 Hf  178 Hf,  175 Yb,  165 Ho,  175 Lu,  176 Lu,  181 Ta,  185 Re,  187 Re,  190 Ir,  193 Ir,  196 Hg,  202 Hg, and  197 Au;    exposing the containers to a neutron source; and    comparing the resulting photon emissions from each of the first and second stable isotopes in each container of the lot to monitor neutron flux variation within the set of samples.    
   
   
       57 . A method for associating each of a plurality of tubes with a particular lot, the method comprising: 
 providing a plurality of containers having a predetermined amount of a first stable isotope composition, M, for identification of the lot, and a known amount of a second stable isotope composition, N, to monitor neutron flux variations, wherein M and N are different and are selected from the group consisting of  36 S;  45 Sc,  50 Cr,  51 V,  55 Mn,  58 Fe,  59 Co,  63 Cu,  75 As,  79 Br,  81 Br,  103 Rh,  107 Ag,  109 Ag,  113 Cd,  114 Cd,  113  In,  115 In,  123 Te,  121 Sb,  123 Sb,  123 Te,  127 I,  133 Cs,  139 La,  141 Pr,  146 Nd,  149 Sm,  152 Sm,  151 Eu,  153 Eu,  152 Gd,  155 Gd,  157 Gd,  159 Tb,  158 Dy,  160 Dy,  161 Dy,  162 Dy,  163 Dy,  164 Dy,  165 Ho,  168 Yb,  169 Tm,  174 Hf,  178 Hf,  175 Yb,  175 Lu,  176 Lu,  181  Ta,  184 Os,  185 Re,  186 W,  187 Re,  190 Ir,  190 Pt,  193 Ir,  196 Hg,  206 Hg, and  197 Au, the containers configured to receive a test sample, wherein the test sample is substantially free of M and N;    exposing the containers to a neutron source; and    detecting the resulting photon emissions at spectra characteristic of M and N, so as to associate each tube of the plurality of tubes with the predetermined quantity of the first stable isotope, for identification of the lot.