# US 12,181,462 B2 · Identification of viable human embryos > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,181,462 B2 - Document ID: US12181462B2 - Status: Granted - Kind: B2 - Application number: 16/937,692 - Filed: 2020-07-24 - Published: 2021-01-28 - Granted: 2024-12-31 - Earliest claimed: 2019-07-25 - Assignees: Overture Life, Inc. - Applicants: Overture Life, Inc. - Inventors: José A. Horcajadas; Hussain A. Abdulla - Web page: https://patentgrep.com/patent/US12181462B2 ## CPC - G01N 33/5005: {involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025)} - G01N 33/5091: {for testing the pathological state of an organism} - G01N 33/689: {related to pregnancy or the gonads} - G01N 2800/36: Gynecology or obstetrics - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract Disclosed herein is a method for assessing the probability of embryo implantation success during the in vitro fertilization process. A method of the disclosure can predict embryo implantation success based on, for example, the amount of specific metabolites present in embryo conditioned culture media. ## Claims ### Claim 1 A method comprising: a) obtaining a sample of culture media in which an embryo has been cultured in vitro; b) i) measuring from the sample an amount of one or more non-pregnancy metabolites associated with a low probability of embryo implantation success, wherein at least one of the non-pregnancy metabolites is indole, leucoline, skatole, 6-hydroxycaproic acid, 1,2,6-hexanetriol, naphthalen-2-amine, 8-hydroxyquinoline, 2,2,6,6-tetramethyl-4-piperidinol, 1,5-naphthalenediamine, 1H-indole-2-carboxylic acid, (E)-dacarbazine, dinitrosopentamethylenetetramine, 5-hydroxyindole-3-acetic acid, 3-amino-5,7-dimethyl-1-adamantanol, 11-aminoundecanoic acid, kynurenine, N-hydroxy-L-tryptophan, 2-(methylamino)-1H-benzo[de]isoquinoline-1,3 (2H)-dione, 5-hydroxy-N-formylkynurenine, or N-lauroylglycine; b) ii) measuring from the sample an amount of one or more pregnancy metabolites associated with a high probability of implantation success, wherein at least one of the pregnancy metabolites is caprolactam, N-cyclohexylformamide, threo-3-Hydroxy-2-methylbutyric acid, sulfoacetic acid, phenylalanine, (2,7-Dimethyloctahydro-1H-cyclopenta[c]pyridin-4-yl)methanol, trans-3-indoleacrylic acid, DL-tryptophan, pantothenic acid, diaveridine, (5-benzyl-3,6-dioxo-2-piperazinyl)acetic acid, or dasytrichone; c) comparing the amount of the one or more non-pregnancy metabolites and the amount of the one or more pregnancy metabolites; and d) transferring the one or more embryos with a high probability of implantation success based on step c) into a subject's uterus. ### Claim 2 (depends on 1) The method of claim 1, wherein one of the pregnancy metabolites is caprolactam. ### Claim 3 (depends on 1) The method of claim 1, wherein one of the pregnancy metabolites is N-cyclohexylformamide. ### Claim 4 (depends on 1) The method of claim 1, wherein one of the pregnancy metabolites is threo-3-hydroxy-2-methylbutyric acid. ### Claim 5 (depends on 1) The method of claim 1, wherein one of the pregnancy metabolites is sulfoacetic acid. ### Claim 6 (depends on 1) The method of claim 1, wherein one of the pregnancy metabolites is (2,7-dimethyloctahydro-1H-cyclopenta[c]pyridin-4-yl)methanol. ### Claim 7 (depends on 1) The method of claim 1, comprising measuring the pregnancy metabolite trans-3-indoleacrylic acid. ### Claim 8 (depends on 1) The method of claim 1, wherein one of the pregnancy metabolites is pantothenic acid. ### Claim 9 (depends on 1) The method of claim 1, wherein one of the pregnancy metabolites is diaveridine. ### Claim 10 (depends on 1) The method of claim 1, wherein one of the pregnancy metabolites is (5-benzyl-3,6-dioxo-2-piperazinyl)acetic acid. ### Claim 11 (depends on 1) The method of claim 1, wherein one of the pregnancy metabolites is dasytrichone. ### Claim 12 (depends on 1) The method of claim 1, comprising measuring the non-pregnancy metabolite indole. ### Claim 13 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is leucoline. ### Claim 14 (depends on 1) The method of claim 1, comprising measuring the non-pregnancy metabolite skatole. ### Claim 15 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is 6-hydroxycaproic acid. ### Claim 16 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is 1,2,6-hexanetriol. ### Claim 17 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is naphthalen-2-amine. ### Claim 18 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is 8-hydroxyquinoline. ### Claim 19 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is 2,2,6,6-tetramethyl-4-piperidinol. ### Claim 20 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is 1,5-naphthalenediamine. ### Claim 21 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is 1H-indole-2-carboxylic acid. ### Claim 22 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is (E)-dacarbazine. ### Claim 23 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is dinitrosopentamethylenetetramine. ### Claim 24 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is 5-hydroxyindole-3-acetic acid. ### Claim 25 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is 3-amino-5,7-dimethyl-1-adamantanol. ### Claim 26 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is 11-aminoundecanoic acid. ### Claim 27 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is kynurenine. ### Claim 28 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is N-hydroxy-L-tryptophan. ### Claim 29 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is 2-(methylamino)-1H-benzo[de]isoquinoline-1,3 (2H)-dione. ### Claim 30 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is 5-hydroxy-N-formylkynurenine. ### Claim 31 (depends on 1) The method of claim 1, wherein one of the non-pregnancy metabolites is N-lauroylglycine. ### Claim 32 (depends on 1) The method of claim 1, comprising measuring the pregnancy metabolite DL-tryptophan. ### Claim 33 (depends on 1) The method of claim 1, wherein one of the pregnancy metabolites is phenylalanine. ### Claim 34 (depends on 1) The method of claim 1, comprising measuring more than one non-pregnancy metabolite and measuring more than one pregnancy metabolite. ### Claim 35 (depends on 34) The method of claim 34, wherein step b) i) comprises measuring each of the non-pregnancy metabolites: indole, leucoline, skatole, 8-hydroxyquinoline, 2,2,6,6-tetramethyl-4-piperidinol, 1,5-naphthalenediamine, 3-amino-5,7-dimethyl-1-adamantanol, 11-aminoundecanoic acid, N-hydroxy-L-tryptophan, and N-lauroylglycine; and step b) ii) comprises measuring each of the pregnancy metabolites: caprolactam, phenylalanine, (2,7-Dimethyloctahydro-1H-cyclopenta[c]pyridin-4-yl)methanol, trans-3-indoleacrylic acid, DL-tryptophan, pantothenic acid, diaveridine, and (5-benzyl-3,6-dioxo-2-piperazinyl)acetic acid. ### Claim 36 (depends on 1) The method of claim 1, wherein the measuring steps comprise a mass spectrometry (MS)-based approach. ### Claim 37 (depends on 36) The method of claim 36, wherein the mass spectrometry (MS)-based approach comprises Fourier transform ion cyctrotron resonance-mass spectrometry (FTICR-MS), Fusion Orbitrap MS, UPLC-coupled Fusion Orbitrap MS, collision-dissociation-tandem MS (CID-MS/MS), higher-energy collision dissociation-coupled tandem MS (HCD-MS/MS), infrared multiphoton dissociation-tandem MS (IRMPD-MS/MS), liquid chromatography triple quadrupole tandem mass spectrometry (LC-QqQ-MS), GC-MS, LC-MS, IC-MS, or capillary electrophoresis-MS (CE-MS). ### Claim 38 (depends on 1) The method of claim 1, wherein the sample of culture media is obtained after an embryo has been cultured in vitro for 0.05 to 8 days. ### Claim 39 (depends on 37) The method of claim 37, wherein step c) comprises calculating a Metabolite Pregnancy Index (MPI) by dividing the total integrated area of mass spectra peaks corresponding to the pregnancy metabolites by the total integrated area of mass spectra peaks corresponding to the non-pregnancy metabolites. ### Claim 40 (depends on 39) The method of claim 39, wherein step d) comprises selecting at least one embryo with an MPI greater than 1.0. ### Claim 41 (depends on 40) The method of claim 40, wherein step d) comprises selecting at least one embryo with an MPI greater than 5. ### Claim 42 (depends on 41) The method of claim 41, wherein step d) comprises selecting at least one embryo with an MPI greater than 10. ### Claim 43 A method comprising: a) obtaining a sample of culture media in which an embryo has been cultured in vitro; b) i) measuring from the sample an amount of one or more non-pregnancy metabolites associated with a low probability of embryo implantation success, wherein at least one of the non-pregnancy metabolites is indole, leucoline, skatole, 6-hydroxycaproic acid, 1,2,6-hexanetriol, naphthalen-2-amine, 8-hydroxyquinoline, 2,2,6,6-tetramethyl-4-piperidinol, 1,5-naphthalenediamine, 1H-indole-2-carboxylic acid, (E)-dacarbazine, dinitrosopentamethylenetetramine, 5-hydroxyindole-3-acetic acid, 3-amino-5,7-dimethyl-1-adamantanol, 11-aminoundecanoic acid, N-hydroxy-L-tryptophan, 2-(methylamino)-1H-benzo[de]isoquinoline-1,3 (2H)-dione, 5-hydroxy-N-formylkynurenine, or N-lauroylglycine; b) ii) measuring from the sample an amount of one or more pregnancy metabolites associated with a high probability of implantation success, wherein at least one of the pregnancy metabolites is caprolactam, N-cyclohexylformamide, threo-3-Hydroxy-2-methylbutyric acid, sulfoacetic acid, phenylalanine, (2,7-Dimethyloctahydro-1H-cyclopenta[c]pyridin-4-yl)methanol, trans-3-indoleacrylic acid, DL-tryptophan, pantothenic acid, diaveridine, (5-benzyl-3,6-dioxo-2-piperazinyl)acetic acid, or dasytrichone; c) comparing the amount of the one or more non-pregnancy metabolites and the amount of the one or more pregnancy metabolites; and d) transferring the one or more embryos with a high probability of implantation success based on step c) into a subject's uterus. ### Claim 44 A method comprising: a) obtaining a sample of culture media in which an embryo has been cultured in vitro; b) i) measuring from the sample an amount of one or more non-pregnancy metabolites associated with a low probability of embryo implantation success, wherein at least one of the non-pregnancy metabolites exhibits a molecular weight of about: 117.06, 129.06, 131.07, 132.08, 134.09, 143.07, 145.05, 157.15, 158.08, 161.05, 182.09, 186.09, 191.06, 195.16, 201.17, 208.08, 220.08, 226.07, 252.07, or 257.20; b) ii) measuring from the sample an amount of one or more pregnancy metabolites associated with a high probability of implantation success, wherein at least one of the pregnancy metabolites exhibits a molecular weight of about: 113.08, 127.10, 132.08, 139.94, 165.08, 183.16, 187.06, 204.09, 219.11, 260.12, 262.09, or 296.10; c) comparing the amount of the one or more non-pregnancy metabolites and the amount of the one or more pregnancy metabolites; and d) transferring the one or more embryos with a high probability of implantation success based on step c) into a subject's uterus.