IP Library › Granted Patent US 12,416,029
Granted Patent B2
US 12,416,029 · App. 17/496,540 · Granted Sep 16, 2025

Compositions and methods for making benzylisoquinoline alkaloids, morphinan alkaloids, thebaine, and derivatives thereof

Inventors: Peter James Facchini (Calgary, CA); Xue Chen (Calgary, CA); Jeffrey C. Colbeck (Davis, CA); Joseph E. Tucker (Strathmore, CA)
Assignee: Antheia Inc.
C12P17/182C12N9/88C12N15/70C12P13/22C12P17/12C12Y114/16002C12Y201/01128
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Quick Facts
Patent No.
US 12,416,029
App. No.
17/496,540
Granted
Sep 16, 2025
Kind
B2
Abstract

Disclosed herein are methods that may be used for the synthesis of benzylisoquinoline alkaloids (“BIAs”) such as alkaloid morphinan. The methods disclosed can be used to produce thebaine, oripavine, codeine, morphine, oxycodone, hydrocodone, oxymorphone, hydromorphone, naltrexone, naloxone, hydroxycodeinone, neopinone, and/or buprenorphine. Compositions and organisms useful for the synthesis of BIAs, including thebaine synthesis polypeptides, purine permeases, and polynucleotides encoding the same, are provided.

Claims (6)

1. A genetically modified cell comprising a recombinant polynucleotide encoding a heterologous thebaine synthase polypeptide wherein the thebaine synthase polypeptide comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO. 6, 31, 32, 80, or 81.

2. The genetically modified cell of claim 1 , wherein the thebaine synthase polypeptide converts salutaridinol-7-O-acetate to thebaine.

3. The genetically modified cell of claim 1 , wherein the thebaine synthase polypeptide comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO. 6, 31, 32, 80, or 81.

4. The genetically modified cell of claim 1 , wherein the thebaine synthase polypeptide comprises the amino acid sequence of SEQ ID NO. 6, 31, 32, 80, or 81.

5. The genetically modified cell of claim 1 , wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO. 19.

6. The genetically modified cell of claim 1 , wherein the cell is a plant cell, fungus, yeast, or bacterium.

Assignments (6)
SECURITY INTEREST Recorded Aug 29, 2025
From: ANTHEIA, INC.
To: HERCULES CAPITAL, INC., AS AGENT
Reel/Frame 072750/0812 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 24, 2022
From: FACCHINI, PETER; CHEN, XUE; TUCKER, JOSEPH
To: WILLOW BIOSCIENCES, INC.
Reel/Frame 059995/0327 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 24, 2022
From: COLBECK, JEFFREY
To: INTREXON CORPORATION
Reel/Frame 059995/0433 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 24, 2022
From: WILLOW BIOSCIENCES, INC.
To: ELESZTO GENETIKA INC.
Reel/Frame 059995/0521 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 24, 2022
From: ELESZTO GENETIKA, INC.
To: ANTHEIA, INC.
Reel/Frame 059995/0716 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 24, 2022
From: PRECIGEN, INC. (FORMERLY INTREXON CORPORATION)
To: ELESZTO GENETIKA, INC.
Reel/Frame 060173/0658 →
Continuity (11)
Continuation 16312776
Provisional Application 62514104 · Jun 2, 2017
Provisional Application 62469006 · Mar 9, 2017
Provisional Application 62438540 · Dec 23, 2016
Provisional Application 62438702 · Dec 23, 2016
Provisional Application 62438601 · Dec 23, 2016
Provisional Application 62438588 · Dec 23, 2016
Provisional Application 62438695 · Dec 23, 2016
Provisional Application 62433431 · Dec 13, 2016
Provisional Application 62355022 · Jun 27, 2016
Related Publication 20220205004A1 · Jun 30, 2022
References Cited (44)
US 6573428B1 · Vodkin et al. · 2003 [cited by applicant]
US 11142780B2 · Facchini et al. · 2021 [cited by applicant]
US 20090156815A1 · Wang et al. · 2009 [cited by applicant]
US 20160208269A1 · Smolke et al. · 2016 [cited by applicant]
WO WO2011058446A2 · 2011 [cited by applicant]
WO WO2015081437A1 · 2015 [cited by applicant]
WO WO2015173590A1 · 2015 [cited by applicant]
WO WO2018005553A1 · 2018 [cited by applicant]
Devos et al., (Proteins: Structure, Function and Genetics, 2000, vol. 41: 98-107. [cited by examiner]
Whisstock et al., (Quarterly Reviews of Biophysics 2003, vol. 36 (3): 307-340. [cited by examiner]
Kisselev L., (Structure, 2002, vol. 10: 8-9. [cited by examiner]
Witkowski et al., (Biochemistry 38:11643-11650, 1999. [cited by examiner]
Fossati et al. PLOS 2015, pp. 1-15. [cited by examiner]
Zulka et al. Planta 2007, 225, pp. 1085-1106. [cited by examiner]
U.S. Office Action dated Apr. 23, 2021 in U.S. Appl. No. 16/312,895. [cited by applicant]
Facchini, P.J. “GenBank Accession No. FE967184” Mar. 31, 2008, [online] [retrieved on Sep. 19, 2017]. Retrieved from the internet: <https://www.ncbi.nlm.nih.gov/nucest/FE967184>. [cited by applicant]
Shitan et al. “Alkaloid Transporters in Plants”, Plant Biotechnology, 31:453-463 (2014). [cited by applicant]
Fossati et al. (2015). “Synthesis of Morphinan Alkaloids in Saccharomyces cerevisiae”, PLoS One 10(4):e0124459. [cited by applicant]
U.S. Non-Final Office Action dated May 20, 2020 in U.S. Appl. No. 16/312,895. [cited by applicant]
Fisinger et al. “Thebaine synthase: A new enzyme in the morphine pathway in Papaver somniferum”, Natural Product Communications, 2:249-253 (2007). [cited by applicant]
Beaudoin et al., “Benzylisoquinoline alkaloid biosynthesis in opium poppy”, Planta, 240: 19-32 (2014). [cited by applicant]
Database EMBL [Online] Jul. 2, 2015 (Jul. 2, 2015), “Papaver somniferum (opium poppy) reticuline epimerase ID-AKO60181; SV 1; linearl; genomic DNA; STD; PLN; 2703 BP.”, retrieved from EBI accession No. Em_CDS: AKO60181. [cited by applicant]
Database Geneseq [Online] Apr. 9, 2015 (Apr. 9, 2015), “Papaver somniferum OMT protein, SEQ ID 543.”, retrieved from EBI accession No. GSP:BBU80692 Database accession No. BBU80692. [cited by applicant]
Winzer et al., “A Papaver somnifarum 10-Gene Cluster for Synthesis of the Anticancer Alkaloid Noscapine”, Science, 336:1704-1708 (2012). [cited by applicant]
Hagel et al., “Dioxygenases catalyze the O-demethylation steps of morphine biosynthesis in opium poppy”, Nature Chemical Biology, 6:273-275 (2010). [cited by applicant]
Morris et al., “Plug-and-Play Benzylisoquinoline Alkaloid Biosynthetic Gene Discovery in Engineered Yeast” in Methods in Enzymology, 144-178 (Elsevier 2016). [cited by applicant]
Galanie et al., “Complete biosynthesis of opioids in yeast”, Science, 349:1095-1100 (2015). [cited by applicant]
Glenn, W.S. et al.' “Recent progress in the metabolic engineering of alkaloids in plant 1-49 systems”, Curr. Opin. Biotechnol. Apr. 2013 (Apr. 2013), vol. 24(2), pp. 354-365. [cited by applicant]
Chen et al., “A pathogenesis-related 10 protein catalyzes the final step in thebaine biosynthesis”, 2018, Nature Chemical Biology, 14:738-743. [cited by applicant]
U.S. Final Office Action dated Nov. 25, 2020 in U.S. Appl. No. 16/312,895. [cited by applicant]
Dastmalchi et al., “Purine Permease-Type Benzylisoquinoline Alkaloid Transporters in Opium Poppy”, 2019, Plant Psychology, 181:916-933. [cited by applicant]
Grothe et al., “Melocular Characterization of the Salutaridinol 7-O-Acetyltransferase Involved in Morphine Biosynthesis in Opium Poppy Papaver somniferum”, 2001, The Journal of Biological Chemistry, 276:30717-30723. [cited by applicant]
Farrow et al. “Stereochemical Inversion of (S)-reticuline by a cytochrome P450 fusion in opium poppy” Nature Chemical Biology, vol. 11, Sep. 2015, p. 728-732. [cited by applicant]
Zulak et al., “Gene transcript and metabolite profiling of elicitor-induced opium poppy cell cultures reveals the coordinate regulation of primary and secondary metabolism,” Planta, 225:1085-1106 (2007). [cited by applicant]
Kisselev L.,“ Polypeptide release factors in prokaryotes and eukaryotes: same function, different structure,” Structure, 2002, vol. 10: 8-9. [cited by applicant]
Witkowski et al., “Conversion of a beta-ketoacyl synthase to a malonyl decarboxylase by replacement of the active-site cysteine with glutamine,” Biochemistry 38:11643-11650, 1999. [cited by applicant]
Whisstock et al., “Prediction of protein function from protein sequence and structure,” Quarterly Reviews of Biophysics 2003, vol. 36 (3): 307-340. [cited by applicant]
Devos et al., “Practical limits of function prediction,” Proteins: Structure, Function and Genetics, 2000, vol. 41: 98-107. [cited by applicant]
Sabarna, “Approaches to isolating a cDNA encoding thebaine synthase of morphine biosynthesis from opium poppy Papaversomniferum L.”, Jun. 22, 2007. [cited by applicant]
Samanani et al., “The role of phloem sieve elements and laticifers in the biosynthesis and accumulation of alkaloids in opium poppy,” Plant Journal, 47:547-563 (2006). [cited by applicant]
International Search Report, issued Dec. 1, 2017, in PCT/US2017/039589. [cited by applicant]
Written Opinion issued Dec. 1, 2017, in PCT/US2017/039589. [cited by applicant]
Choe et al., “Genetic and chemical components analysis of Papaver setigerum naturalized in Korea,” Forensic Science International, 222:387-393 (2012). [cited by applicant]
Facchini et al., “Developmental and inducible accumulation of gene transcripts involved in alkaloid biosynthesis in opium poppy,” Phytochemi, 64:177-186 (2003). [cited by applicant]