US 2014029851 1A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2014/0298511 A1 Lewis et al. (43) Pub. Date: Oct. 2, 2014
(54) BREEDING, PRODUCTION, PROCESSING Publication Classification AND USE OF MEDICAL CANNABS (71) Applicant: BIOTECH INSTITUTE, LLC, (51) Int. Cl. Westlake Village, CA (US) A61E36/85 (2006.01) (52) U.S. Cl. (72) Inventors: Mark Anthony Lewis, Westlake Village, CPC ...... A61K 36/185 (2013.01) CA (US); Michael Dane Backes, Westlake Village, CA (US) USPC ...... 800/260; 424/725; 424/778; 435/410; 8OO/298 (73) Assignee: BIOTECH INSTITUTE, LLC, Westlake Village, CA (US) (21) Appl. No.: 14/216,650 (57) ABSTRACT (22) Filed: Mar 17, 2014 Related U.S. Application Data The invention provides compositions and methods for the (60) Provisional application No. 61/801,528, filed on Mar. breeding, production, processing and use of medical can 15, 2013. nabis. Patent Application Publication Oct. 2, 2014 Sheet 1 of 6 US 2014/029851 1 A1
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BREEDING, PRODUCTION, PROCESSING 0007 Some of the negative or undesirable side effects AND USE OF MEDICAL CANNABS from using available cannabis varieties for medical purposes are related to the plant's content of the chemical A-tetrahy CROSS-REFERENCE TO RELATED drocannabinol (THC). A major hurdle to the more wide APPLICATIONS spread acceptance of cannabis and its legalization is that the land races and commercially available cannabis genotypes 0001. This application claims the benefit of U.S. provi (of drug varieties) contain relatively high concentrations of sional application No. 61/801,528 filed on Mar. 15, 2013 THC. There is a real need for cannabis varieties for potential which is hereby incorporated by reference in its entirety medical use that produce modulated THC concentrations and including all descriptions, references, figures, and claims for varying concentrations of other pharmacologically active all purposes. substances that reduce the negative side effects of THC and increase the medical benefits realized from its use. The inven FIELD OF THE INVENTION tions described herein meet that long-felt need. 0002 The invention relates to cannabis plants for medical use, compositions and methods for making and using said SUMMARY OF THE INVENTION cannabis plants and compositions derived thereof. 0008 According to the methods and compositions of the present invention, plants, plant parts, plant tissues and plant BACKGROUND OF THE INVENTION cells are produced to contain pentyl, propyl, C-4, C-1 and monomethylether constituents of cannabinoid families, 0003 Cannabis, more commonly known as marijuana, is including but not limited to acidic and neutral forms of the a genus of flowering plants that includes at least three species, cannabigerol, cannabichromene, cannabidiol, delta-9-tet Cannabis sativa, Cannabis indica, and Cannabis ruderalis. rahydrohydrocannabinol, delta-8-tetrahydrohydrocannab The use of cannabis for social and medical purposes has been inol, cannabielsoin, cannabinol and cannabinodiol cannab known for almost of all humanity’s recorded history. Can inoid classes; and, cis and trans terpenoids, including but not nabis is most commonly administered via inhalation or con limited to myrcene, limonene, linalool, Ocimene, beta Sumption of marijuana-infused food and drink. pinene, alpha-pinene, beta-caryophyllene, alpha-caryophyl 0004 Since 1972 marijuana has been classified as a lene, delta-3-carene, gamma-bisabolene, alpha-farnesene, Schedule I drug under the U.S. Controlled Substances Act beta-fenchol, guajol, alpha-guaiene, terpinolene, beta-eudes because the U.S. Federal Government considers it to have “no mol, alpha-bergamotene, epi-alpha-bisabolol and caryophyl accepted medical use. In stark contrast to this position, 18 of lene oxide ranging from 0.1% of dry weight of inflorescences, the 50 U.S. states and the District of Columbia have recog plant parts, plant tissues and plant cells to 35% of inflores nized the medical benefits of cannabis and have decriminal cences and/or 95% of plant parts, plant parts, plant tissues and ized its medical use. The 18 U.S. states where medical mari plant cells. juana has been decriminalized as of the filing date of the 0009. The present invention provides medical cannabis present application are as follows: Alaska, Arizona, Califor plants, plant parts, plant tissues and plant cells which provide nia, Colorado, Connecticut, Delaware, Hawaii, Maine, Mas a way to deliver a consistent and more medicinally tolerable sachusetts, Michigan, Montana, Nevada, New Jersey, New and effective ratio of cannabinoids (“CBD) to patients (e.g., Mexico, Oregon, Rhode Island, Vermont and Washington.
2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, BRIEF DESCRIPTION OF THE DRAWINGS 2.9%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, (0018 FIG. 1 illustrates a system 100 for feedback-based 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, cultivation of the herbal medicines described herein. 19.0%, 20.0%, 21.0%, 22.0%, 23.0%, 24.0%, 25.0%, 26.0%, 0019 FIG. 2 Illustrates an exemplary and non-limiting 27.0%, 28.0%, 29.0%, 30.0%, 31.0%, 32.0%, 33.0%, 34.0%, embodiment of the environment management system 104. 35.0%, 36.0%, 37.0%, 38.0%, 39.0%, 40.0%, 41.0%, 42.0%, 0020 FIG.3—Illustrates sensor-to-PC communication, a 43.0%, 44.0%, 45.0%, 46.0%, 47.0%, 48.0%, 49.0%, 50.0%, wireless mesh network of sensors can be employed that feed 51.0%, 52.0%, 53.0%, 54.0%, 55.0%, 56.0%, 57.0%, 58.0%, back to a centralized pc system. 59.0%, 60.0%, 61.0%, 62.0%, 63.0%, 64.0%, 65.0%, 66.0%, 0021 FIG. 4 Illustrates multiplexed cannabis medi 67.0%, 68.0%, 69.0%, 70.0%, 71.0%, 72.0%, 73.0%, 74.0%, cines. 75.0%, 76.0%, 77.0%, 78.0%, 79.0%, 80.0%, 81.0%, 82.0%, 0022 FIG.5 Example of one manifestation of the vapor 83.0%, 84.0%, 85.0%. 86.0%, 87.0%, 88.0%, 89.0%, 90.0%, izer device. 91.0%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99.5%, 0023 FIG. 6—Example of a dosing strip. 99.8%, 99.9% and 100%. DETAILED DESCRIPTION OF THE INVENTION 0013. In some embodiments, the present invention pro vides Medical Cannabis plants, plant parts, tissues and cells 0024 All publications, patents and patent applications, having a THC:CBD ratio of 8:1 and approaching 1:1. By including any drawings and appendices, are herein incorpo rated by reference to the same extent as if each individual comparison, the THC:CBD ratio of the currently available publication or patent application was specifically and indi cannabis varieties is 20:1 and approaches 25:1, 30:1, 35:1, vidually indicated to be incorporated by reference. 40:1 and higher THC to CBD ratios. Thus, the medical can 0025. The following description includes information that nabis plants, plant parts, plant tissues and plant cells of the may be useful in understanding the present invention. It is not present invention will have a THC ratio selected from the an admission that any of the information provided herein is group consisting of 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and 1:1. prior art or relevant to the presently claimed inventions, or 0014. The present invention provides Classes of Cannabis that any publication specifically or implicitly referenced is Varieties developed by selection from landraces of mixed prior art. cannabis genotypes and resulting from further breeding, wherein these Classes of Cannabis Varieties can provide DEFINITIONS useful patient treatment and also are used as breeding mate 0026. As used herein, the verb “comprise' as is used in this rial to develop Medical Cannabis plants and varieties accord description and in the claims and its conjugations are used in ing to the present invention. its non-limiting sense to mean that items following the word 0015 The present invention provides Medical Cannabis are included, but items not specifically mentioned are not plants and varieties with increased organoleptic appeal as a excluded. result of having specified, predetermined terpene and sesquit 0027. The invention provides cannabis plants. As used erpene profiles and content. In some embodiments of the herein, the term “plant” refers to plants in the genus of Can present invention, the increased organoleptic appeal of the nabis and plants derived thereof. Medical Cannabis is inherited in-whole or in-part as a result 0028. The invention provides plant parts. As used herein, of using the Classes of Cannabis Varieties in the breeding the term “plant part” refers to any part of a plant including but program to develop the Medical Cannabis plants. For, not limited to the embryo, shoot, root, stem, seed, stipule, example, in some embodiments, Classes of Cannabis Variet leaf petal, flower bud, flower, ovule, bract, trichome, branch, ies with specific terpene and sesquiterpene profiles and con petiole, internode, bark, pubescence, tiller, rhizome, frond, tent are bred with certain cannabis varieties with specific blade, ovule, pollen, stamen, and the like. The two main parts CBD profiles and content to develop Medical Cannabis Vari of plants grown in Some sort of media, Such as soil or Ver eties with the desired combined attributes of the two types of miculite, are often referred to as the “above-ground' part, also cannabis plants. often referred to as the “shoots', and the “below-ground part, also often referred to as the “roots”. 0016. The present invention also provides methods to 0029. The term “a” or “an refers to one or more of that determine higher THC adequate to down-regulate the entire entity; for example, "a gene' refers to one or more genes or at CB system. This method uses the down-regulation as least one gene. As such, the terms “a” (or “an”), “one or more' therapy for hyper-endocannabinoid systems and to help and “at least one' are used interchangeably herein. In addi increase the therapeutic margin. Additionally, the present tion, reference to “an element' by the indefinite article “a” or invention provides for a potential role of dosage and its influ “an does not exclude the possibility that more than one of the ence on biosynthesis and build-up of cholesterol; a healthy elements is present, unless the context clearly requires that means of Supplementing the endocannabinoid system when there is one and only one of the elements. consuming an ultra low-cholesterol diet. 0030. As used herein, a “landrace' refers to a local variety 0017. The present invention also provides methods for of a domesticated plant species which has developed largely determining the terpene profiles at which dosages are Suit by natural processes, by adaptation to the natural and cultural able for outcomes related to mood elevation and/or sedation environment in which it lives. The development of a landrace (i.e., high limonene for energy, high myrcene for sleep aid, may also involve some selection by humans but it differs from etc.). Furthermore, according to the present invention, terpe a formal breed which has been selectively bred deliberately to nes Such as beta-caryophyllene are used in pain studies (anti conform to a particular formal, purebred Standard of traits. inflammatory via PGE) and linalool is used for anxiety (anti 0031. The International Code of Zoological Nomencla anxiety and sedative). ture defines rank, in the nomenclatural sense, as the level, for US 2014/029851 1 A1 Oct. 2, 2014 nomenclatural purposes, of a taxon in a taxonomic hierarchy applied (artificially or naturally) to the ovule (stigma) of a (e.g., all families are for nomenclatural purposes at the same flower on another plant. Backcrossing is a process in which a rank, which lies between superfamily and subfamily). While breeder repeatedly crosses hybrid progeny, for example a first somewhat arbitrary, there are seven main ranks defined by the generation hybrid (F1), back to one of the parents of the international nomenclature codes: kingdom, phylum/divi hybrid progeny. Backcrossing can be used to introduce one or Sion, class, order, family, genus, and species. more single locus conversions from one genetic background 0032. The invention provides plant cultivars. As used into another. herein, the term “cultivar” means a group of similar plants 0039. The invention provides donor plants and recipient that by structural features and performance (i.e., morphologi plants. As used herein, “donor plants' refer to the parents of a cal and physiological characteristics) can be identified from variety which contains the gene or trait of interest which is other varieties within the same species. Furthermore, the term desired to be introduced into a second variety (e.g., “recipient “cultivar” variously refers to a variety, strain or race of plant plants'). that has been produced by horticultural or agronomic tech 0040. In some embodiments, the present invention pro niques and is not normally found in wild populations. The vides methods for obtaining plant genotypes comprising terms cultivar, variety, strain and race are often used inter recombinant genes. As used herein, the term 'genotype changeably by plant breeders, agronomists and farmers. refers to the genetic makeup of an individual cell, cell culture, 0033. The term “variety' as used herein has identical tissue, organism (e.g., a plant), or group of organisms. meaning to the corresponding definition in the International 0041. In some embodiments, the present invention pro Convention for the Protection of New Varieties of Plants vides homozygotes. As used herein, the term "homozygote (UPOV treaty), of Dec. 2, 1961, as Revised at Geneva on Nov. refers to an individual cell or plant having the same alleles at 10, 1972, on Oct. 23, 1978, and on Mar. 19, 1991. Thus, one or more loci. “variety' means a plant grouping within a single botanical 0042. In some embodiments, the present invention pro taxon of the lowest known rank, which grouping, irrespective vides homozygous plants. As used herein, the term "homozy of whether the conditions for the grant of a breeder's right are gous” refers to the presence of identical alleles at one or more fully met, can be i) defined by the expression of the charac loci in homologous chromosomal segments. teristics resulting from a given genotype or combination of 0043. In some embodiments, the present invention pro genotypes, ii) distinguished from any other plant grouping by vides hemizygotes. As used herein, the term “hemizygotes' the expression of at least one of the said characteristics and iii) or “hemizygous' refers to a cell, tissue, organism or plant in considered as a unit with regard to its suitability for being which a gene is present only once in a genotype, as a gene in propagated unchanged. a haploid cell or organism, a sex-linked gene in the heteroga 0034. As used herein, the term “inbreeding refers to the metic sex, or a gene in a segment of chromosome in a diploid production of offspring via the mating between relatives. The cell or organism where its partner segment has been deleted. plants resulting from the inbreeding process are referred to 0044. In some embodiments, the present invention pro herein as "inbred plants' or “inbreds.” vides heterozygotes. As used herein, the terms "heterozy 0035. The term single allele converted plantas used herein gote' and "heterozygous' refer to a diploid or polyploid refers to those plants which are developed by a plant breeding individual cell or plant having different alleles (forms of a technique called backcrossing wherein essentially all of the given gene) present at least at one locus. In some embodi desired morphological and physiological characteristics of an ments, the cell or organism is heterozygous for the gene of inbred are recovered in addition to the single allele transferred interest which is under control of the synthetic regulatory into the inbred via the backcrossing technique. element. 0036. The invention provides samples. As used herein, the 0045. The invention provides methods for obtaining plant term 'sample' includes a sample from a plant, a plant part, a lines comprising recombinant genes. As used herein, the term plant cell, or from a transmission vector, or a soil, water or air “line' is used broadly to include, but is not limited to, a group sample. of plants Vegetatively propagated from a single parent plant, 0037. The invention provides offspring. As used herein, via tissue culture techniques or a group of inbred plants which the term "offspring refers to any plant resulting as progeny are genetically very similar due to descent from a common from a vegetative or sexual reproduction from one or more parent(s). A plant is said to “belong to aparticular line if it (a) parent plants or descendants thereof. For instance an off is a primary transformant (TO) plant regenerated from mate spring plant may be obtained by cloning or selfing of a parent rial of that line; (b) has a pedigree comprised of a TO plant of plant or by crossing two parent plants and include selfings as that line; or (c) is genetically very similar due to common well as the F1 or F2 or still further generations. An F1 is a ancestry (e.g., via inbreeding or selfing). In this context, the first-generation offspring produced from parents at least one term "pedigree denotes the lineage of a plant, e.g. in terms of of which is used for the first time as donor of a trait, while the sexual crosses affected Such that a gene or a combination offspring of second generation (F2) or Subsequent genera of genes, in heterozygous (hemizygous) or homozygous con tions (F3, F4, etc.) are specimens produced from selfings of dition, imparts a desired trait to the plant. F1's F2s etc. An F1 may thus be (and usually is) a hybrid 0046. The invention provides open-pollinated popula resulting from a cross between two true breeding parents tions. As used herein, the terms "open-pollinated population' (true-breeding is homozygous for a trait), whilean F2 may be or “open-pollinated variety” refer to plants normally capable (and usually is) an offspring resulting from self-pollination of of at least some cross-fertilization, selected to a standard, that said F1 hybrids. may show variation but that also have one or more genotypic 0038. The invention provides methods for crossing a first or phenotypic characteristics by which the population or the plant with a second plant. As used herein, the term "cross’. variety can be differentiated from others. A hybrid, which has “crossing”, “cross pollination' or “cross-breeding refer to no barriers to cross-pollination, is an open-pollinated popu the process by which the pollen of one flower on one plant is lation or an open-pollinated variety. US 2014/029851 1 A1 Oct. 2, 2014
0047. The invention provides self-pollination populations. pistillate structures on individual flowers, whereas monoe As used herein, the term “self-crossing”, “self pollinated or cious plants bear male and female flowers at different loca “self-pollination” means the pollen of one flower on one plant tions on the same plant. is applied (artificially or naturally) to the ovule (stigma) of the same or a different flower on the same plant. 0056 Cannabis is diploid, having a chromosome comple 0048. The invention provides ovules and pollens of plants. ment of 2n=20, although polyploid individuals have been As used herein when discussing plants, the term “ovule' artificially produced. The first genome sequence of Cannabis, refers to the female gametophyte, whereas the term “pollen which is estimated to be 820 Mb in size, was published in means the male gametophyte. 2011 by a team of Canadian scientists. 0049. The invention provides plant tissue. As used herein, 0057 All known strains of Cannabis are wind-pollinated the term “plant tissue' refers to any part of a plant. Examples and the fruit is an achene. Most strains of Cannabis are short of plant organs include, but are not limited to the leaf stem, day plants, with the possible exception of C. sativa Subsp. root, tuber, seed, branch, pubescence, nodule, leaf axil, sativa var. spontanea (C. ruderalis), which is commonly flower, pollen, stamen, pistil, petal, peduncle, stalk, Stigma, described as “auto-flowering and may be day-neutral. style, bract, fruit, trunk, carpel, sepal, anther, ovule, pedicel, needle, cone, rhizome, stolon, shoot, pericarp, endosperm, 0058. The genus Cannabis was formerly placed in the placenta, berry, stamen, and leaf sheath. Nettle (Urticaceae) or Mulberry (Moraceae) family, and later, 0050. The invention provides methods for obtaining plants along with the Humulus genus (hops), in a separate family, the comprising recombinant genes through transformation. As Hemp family (Cannabaceae sensu stricto). Recent phyloge used herein, the term “transformation” refers to the transfer of netic studies based on cpDNA restriction site analysis and nucleic acid (i.e., a nucleotide polymer) into a cell. As used gene sequencing strongly suggest that the Cannabaceae sensu herein, the term “genetic transformation” refers to the transfer stricto arose from within the former Celtidaceae family, and and incorporation of DNA, especially recombinant DNA, that the two families should be merged to form a single into a cell. monophyletic family, the Cannabaceae sensulato. 0051. The invention provides transformants comprising 0059 Cannabis plants produce a unique family of ter recombinant genes. As used herein, the term “transformant' peno-phenolic compounds called cannabinoids. Cannab refers to a cell, tissue or organism that has undergone trans inoids, terpenoids, and other compounds are secreted by glan formation. The original transformant is designated as “TO” or dular trichomes that occur most abundantly on the floral “To Selfing the TO produces a first transformed generation calyxes and bracts of female plants. As a drug it usually comes designated as “T1 or “T” in the form of dried flower buds (marijuana), resin (hashish), 0052. In some embodiments, the present invention pro or various extracts collectively known as hashish oil. There vides plant varieties comprising the recombinant genes. As are at least 483 identifiable chemical constituents known to used herein, the term “variety” refers to a subdivision of a exist in the cannabis plant (Rudolf Brenneisen, 2007, Chem species, consisting of a group of individuals within the spe istry and Analysis of Phytocannabinoids and Other Cannabis cies that are distinct in form or function from other similar Constituents. In Marijuana and the Cannabinoids, ElSohly, arrays of individuals. ed., incorporated herein by reference) and at least 85 different 0053. In some embodiments, the present invention pro cannabinoids have been isolated from the plant (El-Alfy, Abir vides organisms with recombinant genes. As used herein, an T. et al., 2010, Antidepressant-like effect of delta-9-tetrahy "organism” refers any life form that has genetic material drocannabinol and other cannabinoids isolated from Can comprising nucleic acids including, but not limited to, nabis sativa L. Pharmacology Biochemistry and Behavior prokaryotes, eukaryotes, and viruses. Organisms of the 95 (4): 434–42; incorporated herein by reference). The two present invention include, for example, plants, animals, fungi, cannabinoids usually produced in greatest abundance are bacteria, and viruses, and cells and parts thereof. cannabidiol (CBD) and/or A-tetrahydrocannabinol (THC). THC is psychoactive while CBD is not. See, ElSohly, ed. Cannabis (Marijuana and the Cannabinoids, Humana Press Inc., 321 0054 Cannabis has long been used for fibre (hemp), for papers, 2007), which is incorporated herein by reference in its seed and seed oils, for medicinal purposes, and as a recre entirety, for a detailed description and literature review on the ational drug. Industrial hemp products are made from Can cannabinoids found in marijuana. nabis plants selected to produce an abundance of fiber. Some 0060. The cannabinoids in cannabis plants include, but are Cannabis strains have been bred to produce minimal levels of not limited to, A-Tetrahydrocannabinol (A-THC), A-Tet THC, the principal psychoactive constituent responsible for rahydrocannabinol (A-THC), Cannabichromene (CBC), the psychoactivity associated with marijuana. Marijuana con Cannabicyclol (CBL), Cannabidiol (CBD), Cannabielsoin sists of the dried flowers of Cannabis plants selectively bred (CBE), Cannabigerol (CBG), Cannabinidiol (CBND), Can to produce high levels of THC and other psychoactive can nabinol (CBN), Cannabitriol (CBT), and their propyl nabinoids. Various extracts including hashish and hash oil are homologs, including, but are not limited to cannabidivarin also produced from the plant. (CBDV), A-Tetrahydrocannabivarin (THCV), cannab 0055 Cannabis is an annual, dioecious, flowering herb. ichromevarin (CBCV), and cannabigerovarin (CBGV). See The leaves are palmately compound or digitate, with Serrate Holley et al. (Constituents of Cannabis sativa L. XI Canna leaflets. Cannabis normally has imperfect flowers, with bidiol and cannabichromene in samples of known geographi staminate"male' and pistillate “female flowers occurring on cal origin, J. Pharm. Sci. 64:892-894, 1975) and DeZeeuw et separate plants. It is not unusual, however, for individual al. (Cannabinoids with a propyl side chain in Cannabis, plants to bear both male and female flowers. Although mono Occurrence and chromatographic behavior, Science 175:778 ecious plants are often referred to as “hermaphrodites, true 779), each of which is herein incorporated by reference in its hermaphrodites (which are less common) bear staminate and entirety for all purposes. US 2014/029851 1 A1 Oct. 2, 2014
0061 Cannabis plants also produce many types of terpe noids, including, but not limited to, limonene, myrcene, a-pinene, linalool, b-caryophylene, caryophyllene oxide, nerolidol and phytol. 0062 U.S. Pat. No. 6,630,507, assigned on the patent face to The United States of America, is directed to methods of treating diseases caused by oxidative stress by administering therapeutically effective amounts of a CBD that has substan tially no binding to the N-methyl-D-aspartate (NMDA) Tetrahydrocannabinol (THC) receptor, wherein the CBD acts as an antioxidant and neuro protectant. 0063. In one embodiment, the present invention provides 0067. As known as delta-9-tetrahydrocannabinol (A9 medical cannabis plants, which are distinct from the tradi THC), THC is the principal psychoactive constituent (or can tional recreational marijuana plants. nabinoid) of the cannabis plant. The initially synthesized and 0.064 As used herein, medical cannabis or Medical accumulated form in plant is THC acid (THCA). Cannabis refer to cannabis plants, lines, varieties and culti 0068 THC has mild to moderate analgesic effects, and vars having a THC oil content based on the dry weight of plant cannabis can be used to treat pain by altering transmitter inflorescences of greater than or equal to 2.0% but less than or release on dorsal root ganglion of the spinal cord and in the equal to 90% (i.e., THC of > 2.0% and s 90%) and having a periaqueductal gray. Other effects include relaxation, alter CBD oil content based on the dry weight of plant inflores ation of visual, auditory, and olfactory senses, fatigue, and appetite stimulation. THC has marked antiemetic properties, cences equal to or greater than 1.5% (i.e., CBD of e 1.5%); and may also reduce aggression in certain Subjects (Hoaken or, alternatively, having a THC:CBD ratio of 8:1 and (2003). “Drugs of abuse and the elicitation of human aggres approaching 1:1 based on the dry weight of plant inflores sive behavior. Addictive Behaviors 28: 1533-1554). CCCCS. 0069. The pharmacological actions of THC result from its 0065. As a result of the present invention, select cannabis partial agonist activity at the cannabinoid receptor CB, varieties can be used as a physician-recommended form of located mainly in the central nervous system, and the CB medicine or herbal therapy without causing any side effects, receptor, mainly expressed in cells of the immune system or with reduced general or specific side effects when com (Pertwee, 2006. “The pharmacology of cannabinoid recep pared to recreational marijuana plants. Methods for adminis tors and their ligands: An overview”. International Journal of tration of medical cannabis include, but are not limited, to Obesity 30: S13-S18.) The psychoactive effects of THC are vapor inhalation, Smoking (e.g., dried buds), drinking, eating primarily mediated by its activation of CB1G-protein coupled extracts, and taking capsules. receptors, which result in a decrease in the concentration of the second messenger molecule cAMP through inhibition of Cannabis Chemistry adenylate cyclase (Elphick et al., 2001, “The neurobiology and evolution of cannabinoid signalling'. Philosophical 0066 Cannabinoids are a class of diverse chemical com Transactions of the Royal Society B: Biological Sciences 356 pounds that activate cannabinoid receptors. Cannabinoids (1407): 381–408.) It is also suggested that THC has an anti produced by plants are called phytocannabinoids, a.k.a., cholinesterase action which may implicate it as a potential natural cannabinoids, herbal cannabinoids, and classical can treatment for Alzheimer's and Myasthenia (Eubanks et al., nabinoids. At least 85 different cannabinoids have been iso 2006, “A Molecular Link Between the Active Component of lated from the cannabis plants (El-Alfy et al., 2010, Antide Marijuana and Alzheimer's Disease Pathology'. Molecular pressant-like effect of delta-9-tetrahydrocannabinol and Pharmaceutics 3 (6): 773-7.) other cannabinoids isolated from Cannabis sativa L., Phar 0070. In the cannabis plant, THC occurs mainly as tet macology Biochemistry and Behavior 95 (4): 434–42; Bren rahydrocannabinolic acid (THCA 2-COOH-THC). Geranyl neisen, Supra) Typical cannabinoids isolated from cannabis pyrophosphate and olivetolic acid react, catalyzed by an plants include, but are not limited to, Tetrahydrocannabinol enzyme to produce cannabigerolic acid, which is cyclized by (THC), Cannabidiol (CBD), CBG (Cannabigerol), CBC the enzyme THC acid synthase to give THCA. Over time, or (Cannabichromene), CBL (Cannabicyclol), CBV (Cannabi when heated, THCA is decarboxylated producing THC. The varin), THCV (Tetrahydrocannabivarin), CBDV (Cannabidi pathway for THCA biosynthesis is similar to that which pro varin), CBCV (Cannabichromevarin), CBGV (Cannabigero duces the bitter acid humulone in hops. See Fellermeier et al., varin), and CBGM (Cannabigerol Monomethyl Ether). In the (1998, “Prenylation of olivetolate by a hemp transferase Cannabis plant, cannabinoids are synthesized and accumu yields cannabigerolic acid, the precursor of tetrahydrocan lated as cannabinoid acids (e.g., cannabidiolic acid (CBDA)). nabinol”. FEBS Letters 427 (2): 283-5); de Meijer et al. I, II, When the herbal product is dried, stored, or heated, the acids III, and IV (I: 2003, Genetics, 163:335-346; II: 2005, decarboxylize gradually or completely into neutral forms Euphytica, 145:189-198: III: 2009, Euphytica, 165:293-311; (e.g., CBDA->CBD). and IV: 2009, Euphytica, 168:95-112.) US 2014/029851 1 A1 Oct. 2, 2014
Non-Limiting Examples of THC Variants Include: -continued 0071
HO O A-Tetrahydrocannabinolic acid B A-Tetrahydrocannabinol A9-THCB-Cs B A-THC-Cs
A-Tetrahydrocannabinolic acid-C4 A and for B A-THCA-C, A-Tetrahydrocannabinol-C4 A and or B A-THC-C4
A-Tetrahydrocannabivarinic acid A A-Tetrahydrocannabivarin A-Tetrahydrocannabiorcol A-THCVA-C. A A-THCV-C. A-THCO-C
A-Tetrahydrocannabiorcolic acid A-Tetrahydrocannabinolic acid A A and for B 9 A-THCOA-C A-THCA-Cs. A A and or B US 2014/029851 1 A1 Oct. 2, 2014
-continued sea, as well as inhibit cancer cell growth (Mechoulam, et al., 2007, "Cannabidiol recent advances”. Chemistry & Biodi versity 4 (8): 1678-1692.) Recent studies have shown canna bidiol to be as effective as atypical antipsychotics in treating schizophrenia (Zuardi et al., 2006, "Cannabidiol, a Cannabis sativa constituent, as an antipsychotic drug Braz. J. Med. Biol. Res. 39 (4): 421-429.). Studies have also shown that it may relieve symptoms of dystonia (Consroe, 1986, “Open (-)-A-trans-(6aR, 10aR)-A-Tetrahydrocannabinol label evaluation of cannabidiol in dystonic movement disor ders”. The International journal of neuroscience 30 (4): 277
A-THC-Cs 282). CBD reduces growth of aggressive human breast cancer cells in vitro and reduces their invasiveness (McAllister et al., 2007, "Cannabidiol as a novel inhibitor of Id-1 gene expres sion in aggressive breast cancer cells'. Mol. Cancer Ther: 6 (11): 2921-7.) 0073 Cannabidiol has shown to decrease activity of the limbic system (de Souza Crippa et al., “Effects of Canna bidiol (CBD) on Regional Cerebral Blood Flow”. Neuropsy (-)-A-trans-(6aR, 10aR)-Tetrahydrocannabinolic acid A chopharmacology 29 (2): 417-426.) and to decrease Social A-THCA-C5. A
isolation induced by THC (Malon et al., “Cannabidiol reverses the reduction in social interaction produced by low dose A9-tetrahydrocannabinol in rats'. Pharmacology Bio chemistry and Behavior 93 (2): 91-96.) It's also shown that Cannabidiol reduces anxiety in social anxiety disorder (Ber gamaschi et al., 2003, "Cannabidiol Reduces the Anxiety Induced by Simulated Public Speaking in Treatment-Naive Social Phobia Patients'. Neuropsychopharmacology 36 (6): (-)-(6aS, 10aR)-A-Tetrahydrocannabinol 1219-1226). Cannabidiol has also been shown as being effec tive in treating an often drug-induced set of neurological (-)-cis-A-THC-Cs movement disorders known as dystonia (Snider et al., 1985, "Beneficial and Adverse Effects of Cannabidiol in a Parkin son Patient with Sinemet-Induced Dystonic Dyskinesia'. OH Neurology, (Suppl 1): 201.) Morgan etal. reported that strains of cannabis which contained higher concentrations of Can nabidiol did not produce short-term memory impairment vs. strains which contained similar concentrations of THC (2010, OH “Impact of cannabidiol on the acute memory and psychoto Cannabidol (CBD) mimetic effects of Smoked cannabis: naturalistic study: natu ralistic study corrected. British Journal of Psychiatry 197 (4): 285-90.) 0072 CBD is a cannabinoid found in cannabis. It is a major constituent of the plant. Cannabidiol has displayed 0074 Cannabidiol has no affinity for CB and CB recep sedative effects in animal tests (Pickens, 1981, “Sedative tors but acts as an indirectantagonist of cannabinoid agonists. activity of cannabis in relation to its delta'-trans-tetrahydro CBD is an antagonist at the putative new cannabinoid recep cannabinol and cannabidiol content”. Br. J. Pharmacol. 72 tor, GPR55. Cannabidiol has also been shown to act as a (4): 649-56). Some research, however, indicates that CBD can 5-HT1A receptor agonist, an action which is involved in its increase alertness, and attenuate the memory-impairing antidepressant, anxiolytic, and neuroprotective effects. Can effect of THC. (Nicholson et al., June 2004, “Effect of Delta nabidiol is also an allosteric modulator at the Mu and Delta 9-tetrahydrocannabinol and cannabidiol on nocturnal sleep opioid receptor sites. and early-morning behavior in young adults' J Clin Psychop 0075 Cannabis produces CBD-carboxylic acid through harmacol 24 (3): 305-13: Morgan et al., 2010, “Impact of the same metabolic pathway as THC, until the last step, where cannabidiol on the acute memory and psychotomimetic CBDA synthase performs catalysis instead of THCA syn effects of smoked cannabis: naturalistic study, The British thase. See Marks et al. (2009, “Identification of candidate Journal of Psychiatry, 197:258-290). It may decrease the rate genes affecting A9-tetrahydrocannabinol biosynthesis in of THC clearance from the body, perhaps by interfering with Cannabis sativa”. Journal of Experimental Botany 60 (13): the metabolism of THC in the liver. Medically, it has been 3715-3726.) and Meijer et al. I, II, III, and IV. Non-limiting shown to relieve convulsion, inflammation, anxiety, and nau examples of CBD variants include: US 2014/029851 1 A1 Oct. 2, 2014
-continued
(-)-Cannabidiol Cannabidiolic acid CBD-Cs CBDA-Cs
Cannabidiol momomethyl ether Cannabidivarinic acid
CBDVA-C
OH
OH Cannabidiol-C4 Cannabigerol (CBG) CBD-C 0076 CBG is a non-psychoactive cannabinoid found in
the Cannabis genus of plants. Cannabigerol is found in higher concentrations in hemp rather than in varieties of Cannabis cultivated for high THC content and their corresponding psy choactive properties. Cannabigerol has been found to act as a high affinity C2-adrenergic receptor agonist, moderate affin ity 5-HT1A receptor antagonist, and low affinity CB recep torantagonist. It also binds to the CB receptor. Cannabigerol has been shown to relieve intraocular pressure, which may be (-)-Cannabidivarin of benefit in the treatment of glaucoma. Non-limiting CBDV-C, examples of CBG variants include:
OH
2
Cannabigerol Cannabidiorcol CBD-C US 2014/029851 1 A1 Oct. 2, 2014
-continued -continued OH OH O
(SN OH
O H
Cannabigerol monomethyl ether Cannabigerovarinic acid A
(E)-CBGM-CSA--5 (E)-CBGVA-C. A
OH O OH (SN OH
2 O Cannabinol (CBN) Cannabinerolic acid A (Z)-CBGA-Cs A 0077 CBN is a psychoactive substance cannabinoid OH found in Cannabis sativa and Cannabis indica/afghanica. It is (SN also a metabolite of tetrahydrocannabinol (THC). CBN acts as a weak agonist of the CB1 and CB2 receptors, with lower affinity in comparison to THC. Non-limiting examples of 2 CBN variants include
Cannabigerovarin (E)-CBGV-C, OH
OH O S O OH Cannabinol CBN-Cs 2
OH Cannabigerolic acid A (E)-CBGA-Cs. A
O Cannabinol-C4 CBN-C
OH
Cannabigerolic acid A monomethyl ether O (E)-CBGAM-Cs. A Cannabivarin US 2014/029851 1 A1 Oct. 2, 2014 10
-continued (RS)
HO O 25 (-)-Cannabichromene Cannabinol-C2 CBC-Cs
CBN-C (RS)
O 25 O OH Cannabiorcol
(-)-Cannabichromenic acid A CBCA-C5 A
OH O (RS)
OH
O
Cannabinolic acid A HO CBNA-CA (-)-Cannabivarichromene, (-)-Cannabichromevarin CBCV-C.
(RS)
CSO Cannabinol methyl ether CBNM-Cs O OH (-)-Cannabichromevarinic 21 acid A O CBCVA-C. A
OH
HO Cannabichromene (CBC) O 0078 CBC bears structural similarity to the other natural Cannabivarin (CBV) cannabinoids, including tetrahydrocannabinol, tetrahydro cannabivarin, cannabidiol, and cannabinol, among others. Evidence has suggested that it may play a role in the anti 0079 Cannabivarin, also known as cannabivarol or CBV. inflammatory and anti-viral effects of cannabis, and may is a non-psychoactive cannabinoid found in minor amounts in contribute to the overall analgesic effects of medical can the hemp plant Cannabis sativa. It is an analog of cannabinol nabis. Non-limiting examples of CBC variants include: (CBN) with the side chain shortened by two methylene US 2014/029851 1 A1 Oct. 2, 2014 11 bridges (—CH2-). CBV is an oxidation product of tetrahy Non-Limiting Examples of CBL Variants Include: drocannabivarin (THCV. THV). 0083
Cannabidivarin (CBDV) HO (+)-(1aS,3aR,8bR.8cR)- Cannabicyclol 0080 CBDV is a non-psychoactive cannabinoid found in CBL-Cs Cannabis. It is a homolog of cannabidiol (CBD), with the side-chain shortened by two methylene bridges (CH2 units). Plants with relatively high levels of CBDV have been reported in feral populations of C. indica (FC. sativa ssp. indica var. kafiristanica) from northwest India, and in hashish from Nepal. O
O H
O OH (+)-(1aS,3aR,8bR.8cR)- Cannabicyclolic acid A CBLA-C5 A Tetrahydrocannabivarin (THCV, THV)
I0081 THCV, or THV is a homologue of tetrahydrocan nabinol (THC) having a propyl (3-carbon) side chain. This terpeno-phenolic compound is found naturally in Cannabis, Sometimes in significant amounts. Plants with elevated levels of propyl cannabinoids (including THCV) have been found in populations of Cannabis sativa L. ssp. indica (Cannabis indica Lam.) from China, India, Nepal. Thailand, Afghani stan, and Pakistan, as well as Southern and western Africa. HO THCV has been shown to be a CB1 receptor antagonist, i.e. it (+)-(1aS,3aR,8bR.8cR)- blocks the effects of THC. Cannabicyclovarin CBLV-C,
OH OH 8 OH
Cannabicyclol (CBL)
0082 Cannabicyclol (CBL) is a non-psychotomimetic O cannabinoid found in the Cannabis species. CBL is a degra Cannabitriol (CBT) dative product like cannabinol. Light converts cannab ichromene to CBL.
US 2014/029851 1 A1 Oct. 2, 2014 14
-continued I0086) Biosynthetic pathway of cannabinoids has been studied. See Meijer et al. I, II, III, and IV (I: 2003, Genetics, 163:335-346; II: 2005, Euphytica, 145:189-198: III: 2009, Euphytica, 165:293-311; and IV: 2009, Euphytica, 168:95 112), each of which is herein incorporated by reference in its entirety for all purposes. According to the current model, phenolic precursors such as geranyl pyrophosphate (GPP) and polyketide, olivetolic acid (OA) are condensed by gera HO O nyl pyrophosphate olivetolate geranyltransferase (GOT) to a form Cannabigerol acid (CBGA). Alternatively, GPP and Cannabiglendol-C3 divarine acid are condensed by GOT to form Cannabigerova rinic acid (CBGVA). CBGA or CBGAV is transformed to (1) OH-iso-HHCV-C. CBC by CBC synthase or CBCV by CBCV synthase; (2) THC by THC synthase or THCV by THCV synthase; or (3) CBD by CBD synthase or CBDV by CBDV synthase. I0087 More details of cannabinoids synthesis and the properties and uses of these cannabinoids are described in Russo (2011, Taming THC. potential cannabis synergy and phytocannabinoid-terpenoid entourage effects, British Jour nal of Pharmacology, 163: 1344-1364), Russo et al. (2006. A H tale of two cannabinoids: the therapeutic rationale for com bining tetrahydrocannabinol and cannabidiol, Medical Dehydrocannabifuran Hypothesis, 2006, 66:234-246), Celia et al. (Impact of can nabidiol on the acute memory and psychotomimetic effects of smoked cannabis: naturalistic study, The British Journal of Psychiatry, 201, 197:285-290), de Mello Schier et al., (Can nabidiol, a cannabis sativa constituent, as an anxiolytic drug, Rev. Bras. Psiquiatr 2012, 34(S1):5104-5117), and Zhor nitsky et al. (Cannabidiol in Humans—the Quest for Thera peutic Targets, Pharmaceuticals, 2012, 5:529-552), each of which is herein incorporated by reference in its entirety for all purposes. H Cannabifuran Terpenes and Terpenoids in Cannabis Plants I0088 Terpenes are a large and diverse class of organic OH compounds, produced by a variety of plants. They are often strong Smelling and thus may have had a protective function. (SN Terpenes are derived biosynthetically from units ofisoprene, which has the molecular formula CHs. The basic molecular O formulae ofterpenes are multiples of that, (CHs), where n is H the number of linked isoprene units. The isoprene units may be linked together “head to tail to form linear chains or they may be arranged to form rings. Non-limiting examples of Cannabigerovarin (CBGV) terpenes include Hemiterpenes, Monoterpenes, Sesquiterpe nes, Diterpenes, Sesterterpenes, Triterpenes, Sesquarterpe nes, Tetraterpenes, Polyterpenes, and Norisoprenoids. I0089 Terpenoids, a.k.a. isoprenoids, are a large and diverse class of naturally occurring organic chemicals similar to terpenes, derived from five-carbon isoprene units assembled and modified in thousands of ways. Most are mul ticyclic structures that differ from one another not only in CBCV (Cannabichromevarin) functional groups but also in their basic carbon skeletons. OH Plant terpenoids are used extensively for their aromatic quali ties. They play a role in traditional herbal remedies and are under investigation for antibacterial, antineoplastic, and other pharmaceutical functions. Well-known terpenoids include O citral, menthol, camphor, Salvinorin A in the plant Salvia H divinorum, and the cannabinoids found in Cannabis. None limiting examples of terpenoids include, Hemiterpenoids, 1 isoprene unit (5 carbons); Monoterpenoids, 2 isoprene units SooCBGM (Cannabigerol Monomethyl Ether) (10 C); Sesquiterpenoids, 3 isoprene units (15 C); Diterpe noids, 4 isoprene units (20C) (e.g. ginkgolides); Sesterterpe noids, 5 isoprene units (25 C); Triterpenoids, 6 isoprene units US 2014/029851 1 A1 Oct. 2, 2014
(30 C) (e.g. sterols); Tetraterpenoids, 8 isoprene units (40 C) Cannabis sativa constituent, as an anxiolytic drug” (Rev (e.g. carotenoids); and Polyterpenoid with a larger number of Bras Psiquiatr. 2012: 34(Sup11):S104-S117) Also evidence isoprene units. has emerged that CBD can ameliorate the memory impair 0090 Terpenoids are mainly synthesized in two metabolic ment caused by THC. See Morgan, Celia JA, et al. “Impact of pathways: mevalonic acid pathway (a.k.a. HMG-CoA reduc cannabidiol on the acute memory and psychotomimetic tase pathway, which takes place in the cytosol) and MEP/ effects of smoked cannabis: naturalistic study.” The British DOXP pathway (a.k.a. The 2-C-methyl-D-erythritol 4-phos Journal of Psychiatry 1974 (2010): 285-290. phate? 1-deoxy-D-xylulose 5-phosphate pathway, non mevalonate pathway, or mevalonic acid-independent (0096. While THC can be responsible for a range of poorly pathway, which takes place in plastids). tolerated side effects, it has considerable medicinal value, 0091 Cannabis plants produce at over 120 different ter including mood elevation/euphoriant, analgesic, anti-emetic, penes at different levels in their trichomes. Age, maturation anti-inflammatory/antioxidant, bronchodilatory, anti-pru and time of day can affect the amount and ratios of terpenes. ritic, and anti-spasmodic effects, etc. While CBD can reduce Climate and weather also affect terpenes flavonoid produc the side effects of THC, CBD delivers a range of its own tion. In addition to many circulatory and muscular effects, medicinal effects including neuroprotective, anti-oxidant/ Some terpenes interact with neurological receptors. A few anti-inflammatory, anticonvulsant, anti-tumor, antibiotic terpenes produced by cannabis plants also bind weakly to against MRSA, etc. See Russo, Ethan B. “Taming THC: Cannabinoid receptors. Some terpenes can alter the perme potential cannabis synergy and phytocannabinoid-terpe ability of cell membranes and allow in either more or less noid entourage effects.” British Journal of Pharmacology THC, while other terpenes can affect serotonin and dopamine 163.7 (2011): 1344-1364. chemistry as neurotransmitters. (0097. However, an ideal ratio for CBD:THC may be chal 0092 Cannabis usually contains a significant amount of a lenging to achieve because of the complex interactions of terpene called beta-caryophyllene (BCP), which contributes both drugs with various receptors. A recent review by two to the aroma and flavor. Besides BCP, the terpenes produced Canadian researchers illustrates just a few of these chal by cannabis plants include, but are not limited to, borneol. lenges, which states that "overall, the human data regarding caryophyllene, cineole? eucalyptol, Delta-3-carene, CBD's potential to reverse the cognitive perturbations and limonene, linalool, myrcene (e.g., B-myrcene), pinene, pulle psychotomimetic symptoms induced by delta-9-THC are dif gone, Sabinene, terpineol, C-pinene, B-caryophyllene, caryo ficult to interpret due to the possibility of a pharmacokinetic phyllene oxide, nerolidol, and phytol. Cannabis terpenes dis interaction between CBD and delta-9-THC (or other mol play unique therapeutic effects that may contribute ecules) following oral/oromucosal administration.” meaningfully to the entourage effects of cannabis-based medicinal extracts. Phytocannabinoid-terpenoid interactions 0098. When ratio of CBD/delta-9-THC is around 8.1, the have been observed which can produce Synergy with respect CBD displays antagonistic effects. When the ration is 1.8, to treatment of pain, inflammation, depression, anxiety, CBD enhanced the effects of delta-9-THC (Zuardi et al. I, addiction, epilepsy, cancer, fungal and bacterial infections Pharmacological interaction between 9-tetrahydrocannab (including methicillin-resistant Staphylococcus aureus), see inol and cannabidiol, two active constituents of Cannabis Russo (2011, Taming THC. potential cannabis synergy and sativa. Cienc. Cult. 1984, 36, 386-394; Zuardi et al. II, Inter phytocannabinoid-terpenoid entourage effects, British Jour action between cannabidiol (CBD) and A(9)-tetrahydrocan nal of Pharmacology, 163: 1344-1364). nabinol (THC): Influence of administration interval and dose 0093 Geranyl pyrophosphate (GPP), which is used by ratio between the cannabinoids. Psychopharmacology (Berl.) cannabis plants to produce cannabinoids, is formed by con 2012, 219, 247-249). Moreover, there is evidence that com densation of dimethylallyl pyrophosphate (DMAPP) and iso bination of CBD (1-10 mg/kg IP over 21 days) with equiva pentenyl pyrophosphate (IPP) via the catalysis of GPP syn lent doses of delta-9-THC increased blood and brain levels of thase. Alternatively, DMAPP and IPP are ligated by FPP the latter, decreased levels of 11-OH-THC and THC-COOH, synthase to produce farnesyl pyrophosphate (FPP), which can and augmented the anxiogenic and locomotor Suppressant be used to produce sesquiterpenoids. Geranyl pyrophosphate effects and social interaction deficits seen with delta-9-THC. (GPP) can also be converted into monoterpenoids by (Klein et al., Cannabidiol potentiates A9-tetrahydrocannab limonene synthase. inol (THC) behavioral effects and alters THC pharmacoki netics during acute and chronic treatment in adolescent rats. Cannabis Plants Psychopharmacology (Berl.) 2011, 218, 443-457.) 0094 Contemporary illicit “recreational marijuana culti (0099 Interestingly, CBD did not change the THC-induced vars have been exclusively bred and selected primarily for decrease in CB receptor binding and none of the treatments their THC acid content, secondarily for their terpenoid aroma altered 5-HT1A binding, Suggesting that pharmacokinetic and flavor chemistry, and rarely for their production of the factors may have played a role. See Zhornitsky, Simon, and other cannabinoid acids, such as CBDA. Stéphane Potvin. “Cannabidiol in Humans—The Quest for 0.095 CBD reduces or ameliorates some undesirable Therapeutic Targets.” Pharmaceuticals 5.5 (2012): 529-552. effects of THC including intoxication, sedation and tachycar 0100 THC is produced primarily by narrow and broad dia, while contributing analgesic, anti-emetic, and anti-carci leafleted drug cannabis varieties. CBD is produced primarily nogenic properties (Russo and Guy, 2006, Medical Hypoth by narrow and broad leafleted fiber cannabis varieties, com eses (2006) 66, 234-246). Patients that have overmedicated monly known as hemp. Where individual plants are harvested with high-THC cannabis can suffer hallucinations and panic and consumed, breeding for drug cannabis favors selection that persist for up to six hours. Subsequent to this paper, for THC content. Where fields of drug landrace cannabis are evidence has emerged that CBD may contribute anti-anxiety cultivated Solely for extraction, cultivars seem to maintain a effects to cannabis varieties with THC. See "Cannabidiol, a significantly higher percentage of cannabinoids other than US 2014/029851 1 A1 Oct. 2, 2014
THC, including CBD. See Clarke, R C et al. “Cannabis: cannabis and without the threat of overdose or adverse reac Evolution and Ethnobotany’ University of California Press tions found when using pure/high THC marijuana. 2013 (in press). 0106 The present invention also relates to variants, 0101 Almost 99% of cannabis sold by dispensaries in mutants and trivial modifications of the seeds, plant parts California for medical purposes contains less than 1% CBD. and/or whole plants of the cannabis plants of the present (personal communication with SC Laboratories and Halent Laboratory, 2013). Interbreeding drug and hemp varieties of invention. Variants, mutants and trivial modifications of the cannabis can produce cultivars that produce both THC and seeds, plants, plant parts, plant cells of the present invention CBD, in amounts that far exceed landrace cannabis drug or can be generated by methods well known and available to one fiber varieties. See Clarke, R C et al. “Cannabis: Evolution skilled in the art, including but not limited to, mutagenesis and Ethnobotany University of California Press 2013 (in (e.g., chemical mutagenesis, radiation mutagenesis, transpo press). Son mutagenesis, insertional mutagenesis, signature tagged 0102 Importantly to the use of medical cannabis, the ter mutagenesis, site-directed mutagenesis, and natural penoid constituents responsible for the appealing aroma and mutagenesis), knock-outs/knock-ins, antisense and RNA flavor constituents found in the marijuana (i.e., illicit drug) interference. For more information of mutagenesis in plants, variety parent are typically lost in these crosses. These aroma Such as agents, protocols, see Acquaah et al. (Principles of and flavor terpenoids are also synergistic with THC and CBD, plant genetics and breeding, Wiley-Blackwell, 2007, ISBN modulating and enhancing the effects of cannabinoids. 1405136464, 9781405136464, which is herein incorporated Myrcene content in drug cannabis can increase sedation. by reference in its entity). Limonene can reduce anxiety. Pinene is stimulating and 0107 The present invention also relates to a mutagenized enhances memory. But to-date, these terpenoid attributes population of the cannabis plants of the present invention, have not been available in high-CBD varieties. See Russo, and methods of using Such populations. In some embodi Ethan B. “Taming THC: potential cannabis synergy and phy ments, the mutagenized population can be used in screening tocannabinoid terpenoid entourage effects.” British Journal for new cannabis lines which comprises one or more or all of of Pharmacology 163.7 (2011): 1344-1364. the morphological, physiological, biological, and/or chemi 0103) The present invention provides medical cannabis cal characteristics of cannabis plants of the present invention. plants. In some embodiments, the CBD level in dried can In some embodiments, the new cannabis plants obtained nabis plants of the present invention is higher compared to from the screening process comprise one or more or all of the that of a dried recreational cannabis plants, such as the Strain morphological, physiological, biological, and/or chemical White Widow. In some embodiments, the THC level in the dried cannabis plants of the present invention is lower com characteristics of cannabis plants of the present invention, pared to that of a dried recreational cannabis plants, such as and one or more additional or different new morphological, the Strain White Widow. The Cannabis Strain White Widow physiological, biological, and/or chemical characteristic. was developed in The Netherlands from a cross between 0108. The mutagenized population of the present inven sativa and indica. The Strain is known for its abundance of tion can be used in Targeting Induced Local Lesions in white trichomes and high potency. White Widow won the Genomes (TILLING) screening method, which combines a Cannabis Cup in 1995 and is publicly available in most standard and efficient technique of mutagenesis with a chemi Amsterdam “coffeeshops.” In some embodiments, the higher cal mutagen (e.g., Ethyl methanesulfonate (EMS)) with a CBD levels in the cannabis plants of the present invention sensitive DNA screening-technique that identifies single base modulate Some/all of one or more commonly known/per mutations (also called point mutations) in a target gene. ceived side effects of THC. In some embodiments, the present Detailed description on methods and compositions on TILL invention also provides medical cannabis plant varieties with ING(R) can be found in Till et al. (Discovery of induced point specific ratios of CBD to THC. The reduction of THC-related mutations in maize genes by TILLING, BMC Plant Biology adverse effects will result in a better tolerated medicine. 2004, 4:12), Weil et al., (TILLING in Grass Species, Plant 0104. The present invention provides medical cannabis Physiology January 2009 vol. 149 no. 1158-164), Comai, L. plants with specific ratios of terpenes/terpenoids to CBD and S. Henikoff (“TILLING: practical single-nucleotide and/or THC. In some embodiments, the ratio confers syner mutation discovery.” Plant J 45(4): 684-94), McCallum et al., gistic interactions between terpenes/terpenoids and CBD or (Nature Biotechnology, 18:455-457, 2000), McCallum et al., THC. In some embodiments, the ratios of the terpenes/terpe (Plant Physiology, 123: 439-442, 2000), Colbert et al., (Plant noids, CBD and/or THC vary between varieties thereby Physiol. 126(2): 480–484, 2001), U.S. Pat. No. 5,994,075, allowing the creation of mixtures of the varieties to achieve a U.S. Patent Application Publication No. 2004/0053236A1, desired level of one or more of these constituents. In some and International Patent Application Publication Nos. WO embodiments, the terpene/terpenoid levels in the cannabis 2005/055704 and WO 2005/048692, each of which is hereby plants of the present invention are increased or decreased incorporated by reference for all purposes. when compared to that of a control cannabis variety. In some 0109 The present invention also provides any composi embodiments, the control cannabis variety is an existing vari tions or any products made from or isolated from the plants of ety. Such as a recreational marijuana plant variety. The the present invention. In some embodiments, the composi increased terpene/terpenoid content of the varieties of the tions/products comprises extract of the plants, wherein the present invention will be pleasing to patients aesthetically and extract contains more than 2% CBD and less than 98% THC. also modify the effects towards either sedation or stimulation. In Some embodiments, the extract contains higher percentage 0105. The present invention also provides cannabis plants of terpenes/terpenoids compared to extract isolated from a having the same organoleptic appeal as recreational mari control cannabis plant variety (e.g., an existing variety, Such juana plant but with the pharmacological benefit of medical as a recreational cannabis plant variety). US 2014/029851 1 A1 Oct. 2, 2014
Methods of Using Cannabis Plants Tourette's Syndrome, Trichotillomania, Viral Hepatitis, Wasting Syndrome, Whiplash, Wittmaack-Ekbom's Syn 0110. The present invention provides methods ofusing the drome, Writers' Cramp, nausea, vomiting, premenstrual Syn cannabis plants or any parts, any compositions, or any chemi drome, unintentional weight loss, insomnia, and lack of appe cals derived from said plants of the present invention. tite, spasticity, painful conditions, especially neurogenic 0111. In some embodiments, the plants can be used for pain, movement disorders, asthma, glaucoma, adrenal dis medical purpose. In some embodiments, the plants can be ease, inflammatory bowel disease, migraines, fibromyalgia, used by patients having a disease. In some embodiments, the and related conditions, multiple Sclerosis, spinal cord inju diseases includes, but are not limited to, Acquired Hypothy ries. It exhibits antispasmodic and muscle-relaxant properties roidism, Acute Gastritis, Agoraphobia, AIDS Related Illness, as well as stimulates appetite. Other studies State that can Alcohol Abuse, Alcoholism, Alopecia Areata, Alzheimer's nabis or cannabinoids may be useful in treating alcohol Disease, Amphetamine Dependency, Amyloidosis, Amyo abuse, amyotrophic lateral Sclerosis, collagen-induced arthri trophic Lateral Sclerosis (ALS), Angina Pectoris, Ankylosis, tis, asthma, atherosclerosis, bipolar disorder, colorectal can Anorexia, Anorexia Nervosa, Anxiety Disorders, any chronic cer, HIV-Associated Sensory Neuropathy, depression, dysto medical symptom that limits major life activities, any nia, epilepsy, digestive diseases, gliomas, hepatitis C, Chronic Medical Symptom that Limits Major Life Activities, Huntington's disease, leukemia, skin tumors, methicillin-re Arteriosclerotic Heart Disease, Arthritis, Arthritis (Rheuma sistant Staphylococcus aureus (MRSA), Parkinson's disease, toid), Arthropathy, gout, Asthma, Attention Deficit Hyperac pruritus, posttraumatic stress disorder (PTSD), psoriasis, tivity Disorder (ADD/ADHD), Autism/Aspergers, Autoim mune Disease, Back Pain, Back Sprain, Bell's Palsy, Bipolar sickle-cell disease, sleep apnea, and anorexia nervosa. Disorder, Brain Tumor, Malignant, Bruxism, Bulimia, 0112. In some embodiments, the plants of the present Cachexia, Cancer, Carpal Tunnel Syndrome, Cerebral Palsy, invention provide one or more medical benefits to a person in Cervical Disk Disease, Cervicobrachial Syndrome, Chemo need without any side effects, or with reduced side effects therapy Chronic Fatigue Syndrome, Chronic Pain, Chronic compared to a traditional recreational marijuana plant variety. renal failure, Cocaine Dependence, Colitis, Conjunctivitis, In some embodiments, the traditional recreational marijuana Constipation, Crohn's Disease, Cystic Fibrosis, Damage to plant variety is the variety White Widow. In some embodi Spinal Cord Nervous Tissue, Darier's Disease, Degenerative ments, the traditional recreational marijuana plant variety Arthritis, Degenerative Arthropathy, Delirium Tremens, Der contains at least 98%, at least 98.5%, at least 99%, at least matomyositis, Diabetes, Diabetic Neuropathy, Diabetic 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least Peripheral Vascular Disease, Diarrhea, Diverticulitis, Dys 99.9%, or 100% THC in the cannabinoid accumulation in the thymic Disorder, Eczema, Emphysema, Emphysema, plant. Endometriosis, Epidermolysis Bullosa, Epididymitis, Epi 0113. In some embodiments, the plants can be used for lepsy, Felty’s Syndrome, Fibromyalgia, Friedreich's Ataxia, non-medical purposes. In some embodiments, the plants are Gastritis, Genital Herpes, Glaucoma, Glioblastoma Multi used for producing food, oil, wax, resin, rope, cloth, pulp. forme, Graves Disease, Cluster Headaches, Migraine Head fiber, nutrition, construction material, plastic and composite aches, Tension Headaches, Hemophilia A, Henoch-Schon materials, paper, jewelry, water and soil purification materi lein Purpura, Hepatitis C, Hereditary Spinal Ataxia, HIV/ als, weed control materials, cultivation materials, textiles, AIDS, Hospice Patients, Huntington's Disease, clothing, biodegradable plastics, body products, health food Hypertension, Hypertension, Hyperventilation, Hypoglyce and biofuel. mia, Impotence, Inflammatory autoimmune-mediated arthri 0114. In some embodiments, the plants of the present tis, Inflammatory Bowel Disease (IBD), Insomnia, Intermit invention can be used to produce new plant varieties. In some tent Explosive Disorder (IED), Intractable Pain, Intractable embodiments, the plants are used to develop new, unique and Vomiting, Lipomatosis, Lou Gehrig's Disease, Lyme Dis superior varieties or hybrids with desired phenotypes. ease, Lymphoma, Major Depression, Malignant Melanoma, 0.115. In some embodiments, selection methods, e.g., Mania, Melorheostosis, Meniere's Disease, Motion Sick molecular marker assisted selection, can be combined with ness, Mucopolysaccharidosis (MPS), Multiple Sclerosis breeding methods to accelerate the process. Additional breed (MS), Muscle Spasms, Muscular Dystrophy, Myeloid Leu ing methods have been known to one of ordinary skill in the kemia, Nail-Patella Syndrome, Nightmares, Obesity, Obses art, e.g., methods discussed in Chahal and Gosal (Principles sive Compulsive Disorder, Opiate Dependence, Osteoarthri and procedures of plant breeding: biotechnological and con tis, Panic Disorder, Parkinson's Disease, Peripheral ventional approaches, CRC Press, 2002, ISBN 084931321X, Neuropathy, Peritoneal Pain, Persistent Insomnia, Porphyria, 9780849313219), Taji et al. (In vitro plant breeding, Rout Post Polio Syndrome (PPS), Post-traumatic arthritis, Post ledge, 2002, ISBN 156022908X, 9781560229087), Richards Traumatic Stress Disorder (PTSD), Premenstrual Syndrome (Plant breeding systems, Taylor & Francis US, 1997, ISBN (PMS), Prostatitis, Psoriasis, Pulmonary Fibrosis, Quadriple 0412574500, 9780412574504), Hayes (Methods of Plant gia, Radiation Therapy, Raynaud's Disease, Reiter's Syn Breeding, Publisher: READ BOOKS, 2007, drome, Restless Legs Syndrome (RLS), Rheumatoid Arthri ISBN 1406737062, 9781406737066), each of which is incor tis, Rheumatoid Arthritis, Rheumatoid Arthritis, Rosacea, porated by reference in its entirety for all purposes. Cannabis Schizoaffective Disorder, Schizophrenia, Scoliosis, Sedative genome has been sequenced recently (Bakel et al., The draft Dependence, Seizures, Senile Dementia, Severe Nausea, genome and transcriptome of Cannabis sativa, Genome Biol Shingles (Herpes Zoster), Sinusitis, Skeletal Muscular Spas ogy, 12(10):R102, 2011). Molecular makers for cannabis ticity, Sleep Apnea, Sleep Disorders, Spasticity, Spinal Steno plants are described in Datwyler et al. (Genetic variation in sis, Sturge-Weber Syndrome (SWS), Stuttering, Tardive Dys hemp and marijuana (Cannabis sativa L.) according to ampli kinesia (TD). Temporomandibular joint disorder (TMJ), fied fragment length polymorphisms, J Forensic Sci. 2006 Tenosynovitis, Terminal Illness, Thyroiditis, Tic Douloureux, March; 51(2):371-5.), Pinarkara et al., (RAPD analysis of Tietze's Syndrome, Tinnitus, Tobacco Dependence, seized marijuana (Cannabis sativa L.) in Turkey, Electronic US 2014/029851 1 A1 Oct. 2, 2014 18
Journal of Biotechnology, 12(1), 2009), Hakki et al., (Inter time (time to harvest), enhanced nutrient content, novel simple sequence repeats separate efficiently hemp from mari growth patterns, flavors or colors, salt, heat, drought and cold juana (Cannabis sativa L.), Electronic Journal of Biotechnol tolerance, and the like. Desired traits also include selectable ogy, 10(4), 2007), Datwyler et al., (Genetic Variation in Hemp marker genes (e.g., genes encoding herbicide or antibiotic and Marijuana (Cannabis sativa L.) According to Amplified resistance used only to facilitate detection or selection of Fragment Length Polymorphisms, J Forensic Sci, March transformed cells), hormone biosynthesis genes leading to 2006, 51(2):371-375), Gilmore et al. (Isolation of microsat the production of a plant hormone (e.g., auxins, gibberellins, ellite markers in Cannabis sativa L. (marijuana), Molecular cytokinins, abscisic acid and ethylene that are used only for Ecology Notes, 3(1):105-107, March 2003), Pacifico et al., selection), or reporter genes (e.g. luciferase, B-glucuronidase, (Genetics and marker-assisted selection of chemotype in chloramphenicol acetyl transferase (CAT, etc.). The recipient Cannabis sativa L.), Molecular Breeding (2006) plant can also be a plant with preferred chemical composi 17:257-268), and Mendoza et al., (Genetic individualization tions, e.g., compositions preferred for medical use or indus of Cannabis sativa by a short tandem repeat multiplex sys trial applications. tem, Anal Bioanal Chem (2009)393:719-726), each of which 0.121. In some embodiments, molecular markers are is herein incorporated by reference in its entirety for all pur designed and made, based on the genome of the plants of the poses. present application. In some embodiments, the molecular 0116. In some embodiments, said method comprises (i) markers are selected from Isozyme Electrophoresis, Restric crossing any one of the plants of the present invention com tion Fragment Length Polymorphisms (RFLPs), Randomly prising the expression cassette as a donor to a recipient plant Amplified Polymorphic DNAs (RAPDs), Arbitrarily Primed line to create a F1 population; (ii) selecting offspring that have Polymerase Chain Reaction (AP-PCR), DNA Amplification expression cassette. Optionally, the offspring can be further Fingerprinting (DAF), Sequence Characterized Amplified selected by testing the expression of the gene of interest. Regions (SCARs). Amplified Fragment Length Polymor 0117. In some embodiments, complete chromosomes of phisms (AFLPs), and Simple Sequence Repeats (SSRs) the donor plant are transferred. For example, the transgenic which are also referred to as Microsatellites, etc. Methods of plant with the expression cassette can serve as a male or developing molecular markers and their applications are female parent in a cross pollination to produce offspring described by Avise (Molecular markers, natural history, and plants, wherein by receiving the transgene from the donor evolution, Publisher: Sinauer Associates, 2004, ISBN plant, the offspring plants have the expression cassette. 0878.930418, 9780878.930418), Srivastava et al. (Plant bio 0118. In a method for producing plants having the expres technology and molecular markers, Publisher: Springer, sion cassette, protoplast fusion can also be used for the trans 2004, ISBN 1402019114,9781402019111), and Vienne (Mo fer of the transgene from a donor plant to a recipient plant. lecular markers in plant genetics and biotechnology, Pub Protoplast fusion is an induced or spontaneous union, such as lisher: Science Publishers, 2003), each of which is incorpo a Somatic hybridization, between two or more protoplasts rated by reference in its entirety for all purposes. (cells in which the cell walls are removed by enzymatic treat 0.122 The molecular markers can be used in molecular ment) to produce a single bi- or multi-nucleate cell. The fused marker assisted breeding. For example, the molecular mark cell that may even be obtained with plant species that cannot ers can be utilized to monitor the transfer of the genetic be interbred in nature is tissue cultured into a hybrid plant material. In some embodiments, the transferred genetic mate exhibiting the desirable combination of traits. More specifi rial is a gene of interest, Such as genes that contribute to one cally, a first protoplast can be obtained from a planthaving the or more favorable agronomic phenotypes when expressed in expression cassette. A second protoplast can be obtained from a plant cell, a plant part, or a plant. a second plant line, optionally from another plant species or I0123 Details of existing cannabis plants varieties and variety, preferably from the same plant species or variety, that breeding methods are described in Potter et al. (2011, World comprises commercially desirable characteristics, such as, Wide Weed: Global Trends in Cannabis Cultivation and Its but not limited to disease resistance, insect resistance, valu Control), Holland (2010, The Pot Book: A Complete Guide to able grain characteristics (e.g., increased seed weight and/or Cannabis, Inner Traditions/Bear & Co, ISBN 1594778981, seed size) etc. The protoplasts are then fused using traditional 9781594778988), Green I (2009, The Cannabis Grow Bible: protoplast fusion procedures, which are known in the art to The Definitive Guide to Growing Marijuana for Recreational produce the cross. and Medical Use, Green Candy Press, 2009, ISBN 0119) Alternatively, embryo rescue may be employed in 1931160589, 9781931160582), Green II (2005, The Can the transfer of the expression cassette from a donor plant to a nabis Breeder's Bible: The Definitive Guide to Marijuana recipient plant. Embryo rescue can be used as a procedure to Genetics, Cannabis Botany and Creating Strains for the Seed isolate embryos from crosses wherein plants fail to produce Market, Green Candy Press, 1931160279, 9781931160278), viable seed. In this process, the fertilized ovary or immature Starks (1990, Marijuana Chemistry: Genetics, Processing & seed of a plant is tissue cultured to create new plants (see Potency, ISBN 0914171399,9780914171393), Clarke (1981, Pierik, 1999, In vitro culture of higher plants, Springer, ISBN Marijuana Botany, an Advanced Study: The Propagation and 079235267x. 9780792352679, which is incorporated herein Breeding of Distinctive Cannabis, Ronin Publishing, ISBN by reference in its entirety). 091417178X, 9780914171782), Short (2004, Cultivating 0120 In some embodiments, the recipient plant is an elite Exceptional Cannabis: An Expert Breeder Shares His line having one or more certain desired traits. Examples of Secrets, ISBN 1936807122, 9781936807123), Cervantes desired traits include but are not limited to those that result in (2004, Marijuana Horticulture: The Indoor/Outdoor Medical increased biomass production, production of specific chemi Grower's Bible, Van Patten Publishing, ISBN 187882323X, cals, increased seed production, improved plant material 9781878823236), Franck et al. (1990, Marijuana Grower's quality, increased seed oil content, etc. Additional examples Guide, Red Eye Press, ISBN 0929349016, 9780929349015), of desired traits includes pest resistance, vigor, development Groteinhermen and Russo (2002, Cannabis and Cannab US 2014/029851 1 A1 Oct. 2, 2014
inoids: Pharmacology, Toxicology, and Therapeutic Poten additional methods will be developed. However, the efficien tial, Psychology Press, ISBN 07890 15080,97807890 15082), cies of each of these indirect or direct methods in introducing Rosenthal (2007. The Big Book of Buds: More Marijuana foreign DNA into plant cells are invariably extremely low, Varieties from the World's Great Seed Breeders, ISBN making it necessary to use some method for selection of only 1936807068,9781936807062), Clarke, R C (Cannabis: Evo those cells that have been transformed, and further, allowing lution and Ethnobotany 2013 (In press)), King, J (Cannabible growth and regeneration into plants of only those cells that Vols 1-3, 2001-2006), and four volumes of Rosenthal's Big have been transformed. Book of Buds series (2001, 2004, 2007, and 2011), each of 0.126 For efficient plant transformation, a selection which is herein incorporated by reference in its entirety for all method must be employed such that whole plants are regen purposes. erated from a single transformed cell and every cell of the transformed plant carries the DNA of interest. These methods Plant Transformation can employ positive selection, whereby a foreign gene is 0.124 Plants of the present invention can be further modi supplied to a plant cell that allows it to utilize a substrate fied by introducing into the plants one or more transgenes present in the medium that it otherwise could not use, such as which when expressed lead to desired phenotypes. The most mannose or xylose (for example, refer U.S. Pat. No. 5,767, common method for the introduction of new genetic material 378; U.S. Pat. No. 5,994,629). More typically, however, nega into a plant genome involves the use of living cells of the tive selection is used because it is more efficient, utilizing bacterial pathogen Agrobacterium tumefaciens to literally selective agents such as herbicides or antibiotics that either inject a piece of DNA, called transfer or T-DNA, into indi kill or inhibit the growth of nontransformed plant cells and vidual plant cells (usually following wounding of the tissue) reducing the possibility of chimeras. Resistance genes that where it is targeted to the plant nucleus for chromosomal are effective against negative selective agents are provided on integration. There are numerous patents governing Agrobac the introduced foreign DNA used for the plant transforma terium mediated transformation and particular DNA delivery tion. For example, one of the most popular selective agents plasmids designed specifically for use with Agrobacterium— used is the antibiotic kanamycin, together with the resistance for example, U.S. Pat. No. 4,536,475, EP0265556, gene neomycin phosphotransferase (nptII), which confers EP0270822, WO8504899, WO8603516, U.S. Pat. No. 5,591, resistance to kanamycin and related antibiotics (see, for 616, EP0604662, EP0672752, WO8603776, WO9209696, example, Messing & Vierra, Gene 19: 259-268 (1982); Bevan WO9419930, WO9967357, U.S. Pat. No. 4,399.216, et al., Nature 304:184-187 (1983)). However, many different WO8303259, U.S. Pat. No. 5,731,179, EP068730, antibiotics and antibiotic resistance genes can be used for WO9516031, U.S. Pat. No. 5,693,512, U.S. Pat. No. 6,051, transformation purposes (refer U.S. Pat. No. 5,034.322, U.S. 757 and EP904362A1. Agrobacterium-mediated plant trans Pat. No. 6,174,724 and U.S. Pat. No. 6.255,560). In addition, formation involves as a first step the placement of DNA several herbicides and herbicide resistance genes have been fragments cloned on plasmids into living Agrobacterium used for transformation purposes, including the bar gene, cells, which are then Subsequently used for transformation which confers resistance to the herbicide phosphinothricin into individual plant cells. Agrobacterium-mediated plant (White et al., Nucl Acids Res 18: 1062 (1990), Spencer et al., transformation is thus an indirect plant transformation Theor Appl Genet. 79: 625-631 (1990), U.S. Pat. No. 4,795, method. Methods of Agrobacterium-mediated plant transfor 855, U.S. Pat. No. 5,378,824 and U.S. Pat. No. 6,107,549). In mation that involve using vectors with no T-DNA are also addition, the dhfr gene, which confers resistance to the anti well known to those skilled in the art and can have applica cancer agent methotrexate, has been used for selection (Bour bility in the present invention. See, for example, U.S. Pat. No. ouis et al., EMBO.J. 2(7): 1099-1 104 (1983). 7,250,554, which utilizes P-DNA instead of T-DNA in the 0127 Genes can be introduced in a site directed fashion transformation vector. using homologous recombination. Homologous recombina 0.125 Direct plant transformation methods using DNA tion permits site specific modifications in endogenous genes have also been reported. The first of these to be reported and thus inherited or acquired mutations may be corrected, historically is electroporation, which utilizes an electrical and/or novel alterations may be engineered into the genome. current applied to a solution containing plant cells (M. E. Homologous recombination and site-directed integration in Fromm et al., Nature, 319, 791 (1986); H. Jones et al., Plant plants are discussed in, for example, U.S. Pat. Nos. 5,451, Mol. Biol., 13, 501 (1989) and H. Yang et al., Plant Cell 513; 5,501,967 and 5,527,695. Reports, 7,421 (1988). Another direct method, called “biolis I0128 Methods of producing transgenic plants are well tic bombardment, uses ultrafine particles, usually tungsten known to those of ordinary skill in the art. Transgenic plants or gold, that are coated with DNA and then sprayed onto the can now be produced by a variety of different transformation surface of a plant tissue with sufficient force to cause the methods including, but not limited to, electroporation; micro particles to penetrate plant cells, including the thick cell wall, injection; microprojectile bombardment, also known as par membrane and nuclear envelope, but without killing at least ticle acceleration or biolistic bombardment; viral-mediated some of them (U.S. Pat. No. 5,204.253, U.S. Pat. No. 5,015, transformation; and Agrobacterium-mediated transforma 580). A third direct method uses fibrous forms of metal or tion. See, for example, U.S. Pat. Nos. 5,405,765; 5.472,869; ceramic consisting of sharp, porous or hollow needle-like 5,538,877; 5,538,880, 5,550,318; 5,641,664; 5,736,369 and projections that literally impale the cells, and also the nuclear 5,736,369; and International Patent Application Publication envelope of cells. Both silicon carbide and aluminum borate Nos. WO/2002/038779 and WO/2009/117555; Lu et al., whiskers have been used for plant transformation (Mizuno et (Plant Cell Reports, 2008, 27:273-278); Watson et al., al., 2004; Petolino et al., 2000; U.S. Pat. No. 5,302,523 US Recombinant DNA, Scientific American Books (1992); Application 20040197909) and also for bacterial and animal Hinchee et al., Bio/Tech. 6:915-922 (1988); McCabe et al., transformation (Kaepler et al., 1992; Raloff, 1990; Wang, Bio/Tech. 6:923-926 (1988); Toriyama et al., Bio/Tech. 1995). There are other methods reported, and undoubtedly, 6:1072-1074 (1988): Fromm et al., Bio/Tech. 8:833-839 US 2014/029851 1 A1 Oct. 2, 2014 20
(1990); Mullins et al., Bio/Tech. 8:833-839 (1990); Hieiet al., 0.134 Open-Pollinated Populations. Plant Molecular Biology 35:205-218 (1997): Ishida et al., 0.135 The improvement of open-pollinated populations of Nature Biotechnology 14:745-750 (1996); Zhang et al., Such crops as rye, many maizes and Sugar beets, herbage Molecular Biotechnology 8:223-231 (1997); Kuet al., Nature grasses, legumes such as alfalfa and clover, and tropical tree Biotechnology 17:76-80 (1999); and, Raineriet al., Bio/Tech. crops such as cacao, coconuts, oil palm and some rubber, 8:33-38 (1990)), each of which is expressly incorporated depends essentially upon changing gene-frequencies towards herein by reference in their entirety. fixation of favorable alleles while maintaining a high (but far 0129 Microprojectile bombardment is also known as par from maximal) degree of heterozygosity. Uniformity in Such ticle acceleration, biolistic bombardment, and the gene gun populations is impossible and trueness-to-type in an open (Biolistic(R) Gene Gun). The gene gun is used to shoot pellets pollinated variety is a statistical feature of the population as a that are coated with genes (e.g., for desired traits) into plant whole, not a characteristic of individual plants. Thus, the seeds or plant tissues in order to get the plant cells to then heterogeneity of open-pollinated populations contrasts with express the new genes. The gene gun uses an actual explosive the homogeneity (or virtually so) of inbred lines, clones and (.22 caliber blank) to propel the material. Compressed air or hybrids. steam may also be used as the propellant. The Biolistic(R) 0.136 Population improvement methods fall naturally into Gene Gun was invented in 1983-1984 at Cornell University two groups, those based on purely phenotypic selection, nor by John Sanford, Edward Wolf, and Nelson Allen. It and its mally called mass selection, and those based on selection with registered trademark are now owned by E. I. du Pont de progeny testing. Interpopulation improvement utilizes the Nemours and Company. Most species of plants have been concept of open breeding populations; allowing genes to flow transformed using this method. from one population to another. Plants in one population 0130 Agrobacterium tumefaciens is a naturally occurring (cultivar, strain, ecotype, or any germplasm source) are bacterium that is capable of inserting its DNA (genetic infor crossed either naturally (e.g., by wind) or by hand or by bees mation) into plants, resulting in a type of injury to the plant (commonly Apis mellifera L. or Megachile rotundata F.) with known as crown gall. Most species of plants can now be plants from other populations. Selection is applied to improve transformed using this method, including cucurbitaceous one (or sometimes both) population(s) by isolating plants species. A transgenic plant formed using Agrobacterium with desirable traits from both sources. transformation methods typically contains a single gene on 0.137 There are basically two primary methods of open one chromosome, although multiple copies are possible. pollinated population improvement. First, there is the situa Such transgenic plants can be referred to as being hemizy tion in which a population is changed en masse by a chosen gous for the added gene. A more accurate name for Such a selection procedure. The outcome is an improved population plant is an independent segregant, because each transformed that is indefinitely propagable by random-mating within itself plant represents a unique T-DNA integration event (U.S. Pat. in isolation. Second, the synthetic variety attains the same end No. 6,156,953). A transgene locus is generally characterized result as population improvement but is not itself propagable by the presence and/or absence of the transgene. A heterozy as Such; it has to be reconstructed from parental lines or gous genotype in which one allele corresponds to the absence clones. These plant breeding procedures for improving open of the transgene is also designated hemizygous (U.S. Pat. No. pollinated populations are well known to those skilled in the 6,008,437). art and comprehensive reviews of breeding procedures rou 0131 General transformation methods, and specific meth tinely used for improving cross-pollinated plants are pro ods for transforming certain plant species (e.g., maize) are vided in numerous texts and articles, including: Allard, Prin described in U.S. Pat. Nos. 4,940,838, 5,464,763, 5,149,645, ciples of Plant Breeding, John Wiley & Sons, Inc. (1960); 5,501,967, 6,265,638, 4,693,976, 5,635,381, 5,731,179, Simmonds, Principles of Crop Improvement, Longman 5,693,512, 6,162,965, 5,693,512, 5,981,840, 6,420,630, Group Limited (1979); Hallauer and Miranda, Quantitative 6,919,494, 6,329,571, 6,215,051, 6,369,298, 5,169,770, Genetics in Maize Breeding, Iowa State University Press 5,376,543, 5,416,011, 5,569,834, 5,824,877, 5,959,179, (1981); and, Jensen, Plant Breeding Methodology, John 5,563,055, and 5,968.830, each of which is incorporated Wiley & Sons, Inc. (1988). herein by reference in its entirety for all purposes. 0.138 Mass Selection. 0132) Non-limiting examples of methods for transforming 0.139. In mass selection, desirable individual plants are cannabis plants and cannabis tissue culture methods are chosen, harvested, and the seed composited without progeny described in Zweger (The Biotechnology of Cannabis sativa, testing to produce the following generation. Since selection is April 2009); MacKinnon (Genetic transformation of Can based on the maternal parent only, and there is no control over nabis sativa Linn: a multi purpose fibre crop, doctoral thesis, pollination, mass selection amounts to a form of random University of Dundee, Scotland, 2003), MacKinnon et al. mating with selection. As stated herein, the purpose of mass (Progress towards transformation of fibre hemp, Scottish selection is to increase the proportion of Superior genotypes Crop Research, 2000), and US 20120311744, each of which in the population. is herein incorporated by reference in its entirety for all pur 0140) Synthetics. poses. The transformation can be physical, chemical and/or 0.141. A synthetic variety is produced by crossing interse biological. a number of genotypes selected for good combining ability in all possible hybrid combinations, with Subsequent mainte Breeding Methods nance of the variety by open pollination. Whether parents are 0.133 Classical breeding methods can be included in the (more or less inbred) seed-propagated lines, as in Some Sugar present invention to introduce one or more recombinant beet and beans (Vicia) or clones, as in herbage grasses, clo expression cassettes of the present invention into other plant vers and alfalfa, makes no difference in principle. Parents are varieties, or other close-related species that are compatible to selected on general combining ability, sometimes by test be crossed with the transgenic plant of the present invention. crosses or toperosses, more generally by polycrosses. Paren US 2014/029851 1 A1 Oct. 2, 2014 tal seedlines may be deliberately inbred (e.g. by selfing or sib Example 1 crossing). However, even if the parents are not deliberately inbred, selection within lines during line maintenance will ensure that Some inbreeding occurs. Clonal parents will, of Development of Classes of Cannabis Varieties course, remain unchanged and highly heterozygous. 0151. One objective of the present invention was to survey 0142. Whether a synthetic can go straight from the paren landraces of cannabis to identify and establish classes of tal seed production plot to the farmer or must first undergo known cannabis plants, wherein the individual plants of the one or two cycles of multiplication depends on seed produc present invention (i.e., cultivars, varieties, strains, races) tion and the scale of demand for seed. In practice, grasses and within a given class of cannabis have certain common physi clovers are generally multiplied once or twice and are thus ological, chemical and/or morphological characteristics. considerably removed from the original synthetic. Thus, according to the present invention, cannabis plants are 0143. While mass selection is sometimes used, progeny grouped into named classes according to their primary/domi testing is generally preferred for polycrosses, because of their nant flavor(s) in order to establish standard cannabis classes operational simplicity and obvious relevance to the objective, of plants herein referred to collectively as Classes of Can namely exploitation of general combining ability in a syn nabis Varieties. In some instances, class names are indicative thetic. of the landrace plant type from which they originated. 0144. The numbers of parental lines or clones that enter a synthetic vary widely. In practice, numbers of parental lines 0152. As explained in greater detail below, individual can range from 10 to several hundred, with 100-200 being the nabis plants were identified, tested and selected within each average. Broad based synthetics formed from 100 or more landrace to form varieties within each cannabis class. Accord clones would be expected to be more stable during seed ing to the present invention, more than one variety of Can multiplication than narrow based synthetics. nabis may have been established within a single cannabis class. As explained further herein, selected candidate Can (0145 Pedigreed Varieties. nabis plants for a specific variety may have been Subjected to 0146 A pedigreed variety is a Superior genotype devel further breeding and selection before being chosen as a Can oped from selection of individual plants out of a segregating nabis variety for a particular class. The final selected varieties population followed by propagation and seed increase of self were designated as Classes of Cannabis Varieties. Therefore, pollinated offspring and careful testing of the genotype over as used herein, Classes of Cannabis Varieties or variety several generations. This is an open pollinated method that Classes or the like each refer to certain cannabis varieties works well with naturally self pollinating species. This originating out of landraces, wherein they were selected method can be used in combination with mass selection in based on certain desirable phenotypical characteristics and variety development. Variations in pedigree and mass selec morphological characteristics for a particular class of Can tion in combination are the most common methods for gen nabis. erating varieties in self pollinated crops. 0147 Hybrids. 0153. The following Table 1 provides the Class color, Class name and Class abbreviations (ABRV) for each Class 0148. A hybrid is an individual plant resulting from a cross of the Classes of Cannabis Varieties provided by the present between parents of differing genotypes. Commercial hybrids invention. are now used extensively in many crops, including corn (maize), Sorghum, Sugarbeet, Sunflower and broccoli. TABLE 1 Hybrids can beformed in a number of different ways, includ ing by crossing two parents directly (single cross hybrids), by Cannabis class name abbreviations. crossing a single cross hybrid with another parent (three-way or triple cross hybrids), or by crossing two different hybrids CLASS CLASS (four-way or double cross hybrids). NAME ABRV 0149 Strictly speaking, most individuals in an out breed GOLD GOD BRONZE BRO, HAZ ing (i.e., open-pollinated) population are hybrids, but the term WHITE WHI is usually reserved for cases in which the parents are indi FUSCIA CHM viduals whose genomes are sufficiently distinct for them to be ORANGE ORA recognized as different species or subspecies. Hybrids may be RED RED fertile or sterile depending on qualitative and/or quantitative YELLOW YEL differences in the genomes of the two parents. Heterosis, or SILVER SIL hybrid vigor, is usually associated with increased heterozy PURPLE PUR gosity that results in increased vigor of growth, Survival, and GRAY GRA JADE JCK, JK fertility of hybrids as compared with the parental lines that BLUE BLU were used to form the hybrid. Maximum heterosis is usually BLACK BLA achieved by crossing two genetically different, highly inbred GREEN GRE lines. MISC MI 0150. This invention is further illustrated by the following examples which should not be construed as limiting. The contents of all references, patents and published patent appli 0154 The following Table 2 provides the variety names cations cited throughout this application, as well as the Fig for each of the Classes of Cannabis Varieties provided by the ures are incorporated herein by reference. present invention. US 2014/029851 1 A1 Oct. 2, 2014 22
TABLE 2
Classes of cannabis varieties
GO13 JK21 CBD2 HAZZ, BLA14 BLA19 PUR24 MI11 G BLA44 SIL4 K2O CBD4 BLU7 BLA1S BLA20 PUR2S MI12 G BLA45 PUR3 JK16 CBDS GRE2 BLA16 BLA21 PUR26 MI13 G BLA46 YEL3 JK14 PUR4 WHI4 BLA17 BLA22 PUR27 MI14 THV1 BLA47 GRE CKS PURS WHIS BLA18 BLA23 PUR28 MI15 TEHV1 BLA48 WHI2 BRO4 PUR6 WHIS BLA19 BLA24 PUR29 MI16 THV1 BLA49 WHI3 ORA3 HAZZ, PUR3O BLA2O BLA2S ISO1 MI17 THV1 BLASO YEL4 ORA2 BLU7 GRE30 BLA21 BLA26 ISO2 MI18 THV1 BLAS1 GRA3 BROS GRE2 WHI6 BLA22 BLA27 ISO3 MI19. THV1 BLAS2 RED2 SIL2 WHI4 GRE31 BLA23 BLA28 ISO4 MI2O THV1 BLA44 CBD3 BLU1 WHIS HAZS BLA24 BLA29 ISOS GRO3 THV1 BLA45 BLA1 BLU2 WHIS HAZ6 BLA2S BLA30 ISO6 GRO4 THV1 BLA46 WHIf SIL3 PUR3O HAZ7 BLA26 BLA31 ISO7 GROS THV1 BLA47 GOD1 BLU3 GRE30 HAZ8 BLA27 BLA32 ISO8 GRO6 THV1 BLA48 GOD2 HAZ3 WHI6 GO14 BLA28 BLA33 ISO9 GRO7 THV2 BLA49 GOD3 HAZ2 GRE31 BLU8 BLA29 BLA34 IS10 GRO8 THIV2 BLASO GOD4 ORA4 HAZS BLU9 BLA3O PURA IS11 GRO9 THV2 BLAS1 GODS GRA2 HAZ6 BLU10 BLA31. PUR8 IS12 GR1O THV2 BLAS2 BRO1 CBD1 HAZ7 BLU11 BLA32 PUR9 IS13 GR11 THV2 WHI1 HAZA. HAZ8 BLU12 BLA33 PUR1O IS14 GR12 THV2 CHM1 GOD6 GO14 BLU13 BLA34 PUR11 IS15 GR13 THV2 ORA1 GOD7 BLU8 BLA2 PURA PUR12 IS16 GR14 THV2 CHM2 GOD8 BLU9 BLA3 PUR8 PUR13 IS1.7 GR1S THV2 RED1 GOD9 BLU10 BLA4 PUR9 PUR 14 MIO1 GR16 THV2 YEL1 GO10 BLU11 BLAS PUR1O PUR1S MIO2 GR17 BLA35 SIL1 GO11 BLU12 BLA6 PUR11 PUR16 MIO3 GR18 BLA36 PUR1 YEL2 BLU13 BLA7 PUR12 PUR17 MIO4 GR19 BLA37 GRA1 CHM3 CBD2 BLA8 PUR13 PUR18 MIOS GR2O BLA38 JCK4 GO12 CBD4 BLA9 PUR14 PUR19 MO6 GR21 BLA39 JCK8 PUR2 CBDS BLA10 PUR1S PUR20 MIO7 GR22 BLA40 JCK7 BLU4 PUR4 BLA11. PUR16 PUR21 MIO8 GR23 BLA41 JK12 BLUS PURS BLA12 BLA17 PUR22 MI09 GR24 BLA42 JK11 BLU6 PUR6 BLA13 BLA18 PUR23 MI1 O GR2S BLA43
0155 The following Table 3 provides the predominant 0160 Top Mix ground up entire top bud (usually top 6" flavors of each Class of Classes of Cannabis Varieties accord- of plant). ing to the present invention. 0.161 Low Mix ground up lower buds (under top canopy). TABLE 3 0162 Standard Mix ground-up combination of lowest, middle and top buds in order to determine an overall average Cannabis class predominant flavors for plant chemotype. Class Class Flavor (0163 Max THC Wt.% total possible amount of delta 9 GOLD Lemon Pine THC that can result from the combustion of THCA (about SILVER Sandalwood Coffee/Spice 70% conversions at best). For a detailed explanation of the BRONZE Hash/Spice? Haze methodology to determine Max THC Wt. '% see Dussy et al. PURPLE Grape? Pine BLUE Blueberry/Pine (2005) Isolation of A-THCA-A from hemp and analytical GREEN Sweet Citrust Cream Skunk aspects concerning the determination of A-THC in cannabis RED Cherry/Sweet products. Forensic Science International 149:3-10. ORANGE MandarinOrange, Lemon YELLOW Spruce/Spice? Pine (0164. As shown further herein, the Classes of Cannabis WHITE Vanilla Varieties of the present invention are used for medical pur GRAY Generic Sweet Amsterdam poses as is or are used in breeding schemes to develop new BLACK Anise Black Licorice cannabis varieties within, between or among the various JACK Sweet Myrcene/Pine classes of Classes of Cannabis Varieties. FUSEHIA Chemy MISC Various GO13 0156 More detailed descriptions of the development and 0.165 Description of Breeding Stock. characteristics of the Classes of Cannabis Varieties of the (0166 Inflorescences were obtained for a land race of Gold present invention are provided below, including for the fol class varieties and seeds from these inflorescences were iso lowing plant traits: lated and put into conditions proper for their germination. The 0157 Sugar Leaf leaves surrounding buds (inflores seeds which germinated grew identically. However, upon cences) and that are covered with glandular trichomes. flower onset, the seedlings were selected for the strongest 0158 Top of Top—apical most inflorescence body (very limonene/Pine-Sol fragrance and narrowed to two pheno tip top of plant). types. Of these, the individual phenotype with the best user 0159 Low of Top lowest auxiliary bud that is part of experience based on testing was selected to create GO13, a apical most inflorescence (bottom bud of tip top of plant). variety classified into the Gold Class. US 2014/029851 1 A1 Oct. 2, 2014
(0167 Hypothesized Genetics. proprietary soil mix A and remain in 18 hours of daylight for 0168 Cannabis indica ssp. afghanica WLD “Purple another 14-21 days. Once root bound, plants are transplanted Afghan'xC. indica ssp. indica var. indochinensis NLD into rigid 3 gallon containers filled with proprietary soil mix “Lemon Thai'xC. indica ssp. kafiristanica NLDA. B. Immediately, the light cycle is altered to 12/12 and flower 0169 Propagation and Vegetative Growth. initiating begins. The plants remain in 12/12 lighting until 0170 Cuttings from GO13 are marked by 3-finger leaflet harvesting. They undergo a propriety nutrient regimen and sets with internode buds asymmetrically located on alternate grow as undisturbed as possible for 60-70 days depending on sides on main shoot. In particular, the internode space of this chemotype analysis. variety tends to be greater than that of other gold class Vari 0180 All sun leaves are removed and plant dismantled to eties and stems harden quicker. Roots nodes appear with 7-10 result in approximately 12" branches covered in inflores days and roots within 10-14 days. The GO13 grows extremely cences and trichomes. The goal in harvesting is to realize that tall and thin with extreme stretching and asymmetrical bud we are actually harvesting trichome heads but not buds. and leaf sets. When root system is not limited or pruned, this Thus, great care is taken not to disturb the trichome heads and variety of gold class varieties exhibits unparalleled vigor and as much of the plant remains intact as possible to promote stretch. Vegetative growth is marked by a deep blue-green even and slow drying. (Munsell ID) hue with lime green thin stalks. Petioles are 0181. Yield Data. marked by purple pointillism increasing on sides exposed to 0182 Yield determined on a per plant basis and deter light and the end closest to palm of the leaflet set. Root bodies mined by specified cultivation techniques employed. In this are typically full and bright white. Stalks radiate a pungent case, indoor Controlled Environment Agriculture (CEA) Smell of body odor or urine. Canopy extremely sparse and technique following the protocol described elsewhere herein. apical dominance can be disrupted easily with removal of Flower onset was initiated with 12/12 day/night at approxi apical meristem. Main stems also exhibit purpling, but inflo mately 12" in vegetative height. Total biomass ~150 g, fin rescences are not purple. ished flowers ~50 g, and/or ~50 g of seed per plant. 0171 Onset of Flowering and Inflorescences. 0183 Potential Uses of this Line. 0172 Leaves are 3 and 5 leaflet patterns with 3 being 0.184 Potential uses of GO13 include but are not limited to predominant and overall decreasing to 1 and to none in the medical applications, as a source for extractions of plant presence of female flowers. constituents and chemicals, for commercial raw materials, 0173 Female flowers are spread out due to the large inter fiber and pulp. node spacing. Upon flower set, buds and Supporting struc 0185. Patient Testimonials/Comments and Visual Obser tures (stems, leaves, etc.) are quickly covered with an Vations. extremely dense field of trichome bodies. Again, this variety 0186 Patients rave about the flavor and oily composition tends to be more densely covered with trichome bodies than by comparison to other Gold class varieties. In fact, besides its parent and other gold class varieties. In fact, the inflores the extremely high potency from its combined cannabinoid/ cences are very dense and have large calyxes covered in terpenoid entourage effects', this line of gold class has been highly resinous glands that exhibit this variety’s distinct noted by patients for being particularly effective for sexual lemon Pine-Sol scent after only 7-10 days. As inflorescences mature, the density compact sets give way to foxtailing and and appetite stimulation. reaching by individual calyxes, resulting in an overall 0187 Palatable CBDA varieties with ideal CBDA:THCA increase in Surface area dedicated to trichome production. In ratio can be developed from GO13 to reduce side-effects particular, the oily character of these flowers set this gold associated with extant recreational cannabis varieties related class apart from its parent and other gold class varieties. to GO13. Additionally reduced THCA varieties can be devel Textures are extremely sticky and fibrous. Stems do not oped that are intended to reduce side-effects from extant break they tear, but remain attached via intense fiber strands. recreational cannabis varieties related to GO 13. (0174 Description of Finished Flower. 0188 Flavor when smoked included distinct citrus and 0175 GO13 consistently produces among the highest mentholated notes. Significant analgesia accompanies its THCA levels of medical cannabis known in California and is deep range of effects, but with little sedation, but the “rising/ often noted for an intense and crushing physical effect com falling physical sensations associated with gold class. Some bined with a Sublime and inspiring mental flight. Aromas of patients have compared its flavor to bergamotorange. Patients lemon peel, NASCAR, and Pine-Sol combine to produce a also remark on the “clarity” of this variety's psychoactivity, pure menthol exhalation when smoked. Noted for excellent with less sedation and disorientation, and with considerable appetite and sexual stimulation often accompanied by unin euphoria. terrupted sleep. 0189 Its aroma has been characterized as a tangy, sharp, (0176 Chemotype Description for Patient. naphthalene aroma with orange notes and a Sweet undertone. 0177 Relative potency: very strong. Headspace Terpenes: Also the range of pharmacologically active terpenoids that pinenes, myrcene, limonene, and linalool. Caryophyllene this variety produces provide a significant "entourage effect” content: high. that accompanies the effects of its THC content. While it 0178. Description of Planting, Harvesting and Processing stimulates appetite, it does not appear to encourage overeat of the Plants. ing. 0179 This variety is asexually propagated via taking cut (0190. Chemical Analysis. tings of shoots and putting them in rock wool cubes. These (0191 Low MW Terpene Analysis (136). Sesquiterpene cubes were presoaked with pH adjusted water and kept warm Analysis (204). Cannabinoid Analysis (>284). (-80°C.). Full trays were covered, left under 18 hours of light 0.192 Mass spectrometry data, chromatography data, and and allowed to root (7-14 days). Upon root onset, the plantlets statistical data related to the chemical analysis are shown in were transplanted into rigid 1 gallon containers filled with a the following Tables 4-6. US 2014/029851 1 A1 Oct. 2, 2014 24
TABLE 4 Chemical analysis of plant samples for GO13. Plant THC wt.% THCA CBD wt.% CBDA CBN THC, CBD Chrono
Location Ave Max wt.% Ave Max wt.% wt.% Ratio ID Sample #: see Chromo ID numbers in last column. Sugar leaf 0.5 4.1 6.O O.1 O.1 O.O O.1 41 WRC-007 Top of top 0.5 9.6 14.7 O.2 O.2 O.1 O.2 48 WRC-008 Low of O.9 11.3 16.9 O.1 O.2 O.1 O.2 56.5 WRC-009 top Top mix O.7 11.2 17.0 O.2 O.2 O.1 O.3 56 WRC-17 Standar 1.O 113 16.8 O.2 O.2 O.1 O.2 56.5 WRC-18 mix Sample #: AS623
Standar O.8 11.3 16.8 O2 O2 O.1 O.1 56.5 mix Sample #: A8623
Standar O.7S 15.51 16.81 0.18 O.25 O.08 ND 61.07 Mix
THCCBDRatio = Maximum THC wt %Maximum CBD wt %. Chromo D = Chromatogram identification number for quantification of cannabinoids, ND = Not determined.
TABLE 5 TABLE 6-continued
Chemical analysis of flower samples for GO13 for Sample #: 2876-7.1. The cannabinoid and terpene profiles, unheated, % by wit, or terpene content by retention time for Sample #: 2876-7.1 THC CBD CBN Cannabinoid Terpenoid Terpene content Profile % by wt Profile % by wt by retention time Average Maximum THCA Average Maximum CBDA Maximum CBGA O.32 22-30 sesquiterpines? CBG O.43 caryophyllenes Total content for unheated flower sample, 9% by wt. CBLA O.10 CBNA <0.03 0.73 ND 18. SS <0.1 ND <0.1 <0.1 CBN <0.03 Total content for unheated flower sample, mg/g
7.3 ND 185.5 <0.1 ND O.6 O.1 Estimated total content for ideally heated flower sample, % by wt. BRO5 (0193 Description of Breeding Stock. ND 12.1 ND ND <0.1 ND <0.1 0194 Inflorescences were obtained for a landrace of Haze and seeds from these inflorescences were isolated and put into ND = Not determined. conditions proper for their germination. Note: 0.195 The seeds which germinated grew identically, being Most of the acid cannabinoids (e.g., THCA) readily converts to their neutrals (THC) upon short and squatty with purple leaves and Sweet scent, with heating, giving different properties, one exception which was tall and stretchy with a savory and musty scent. There was absolutely no Sweetness in the Smell TABLE 6 of BRO5. Testing proved that its effects were the most enjoy able and virtually mycrene free. The lack of mycrene and The cannabinoid and terpene profiles, unheated, % by wit, or terpene presence of pinene and limonene is quite rare and sets this content by retention time for Sample #: 2876-7.1 variety apart from most cannabis varieties. Cannabinoid Terpenoid Terpene content 0196. Upon flower onset, the seedlings were selected for Profile % by wt Profile % by wt by retention time being short and squatty with purple leaves and berry scent to THCA 18.55 terpinolene O.11 5.91 alpha-pinene create BRO5, a variety classified into the Gold Class. A-THC 0.73 myrcene O.86 6.4 camphene (0197) Hypothesized Genetics. A8-THC <0.03 Limonene <0.01 7.35 beta-pinene (0198 “NL#5x Hazex inbred Thai” THCA-C4 0.04 Linalool O.18 7.91 myrcene THCVA 0.12 Caryophylene <0.01 9.53 carpenesterpins 0199 Propagation and Vegetative Growth. Oxide 0200 Cuttings from BRO5 are marked by 9-finger very THCV <0.03 Beta- O.31 9.34 alpha-limonene thin leaflet sets with internode buds asymmetrically located Caryophyllene CBDA 0.06 Phytol <0.01 10.15 ocimene on alternate sides on main shoot. In particular, the internode CBD <0.03 alpha-pinene O.O7 11.66 terpinolene space of this variety tends to be extremely large. Stems are CBCA O.O3 12.39 linalool tall, frail and stretchy. Cuttings roots appear within 10-14 CBC <0.03 16.41 terpineol days. The BRO5 grows tall and stretchy with flimsy stems. It possesses the classic narrow-leafleted morphology associated US 2014/029851 1 A1 Oct. 2, 2014
with 1970’s Haze cultivars that were inherited from HaZe’s case, indoor CEA technique following the protocol described tropical drug cannabis parents, including Colombian and elsewhere herein. Flower onset was initiated with 12/12 day/ That varieties. night at approximately 16" in vegetative height. Organic mix 0201 BRO5 grows with asymmetrical bud and leaf sets. of Soil in fabric pots, a regimen of nutrients following stan Vegetative growth is marked by a lightened green (Munsell dard NPK feeding schedules and addition of proprietary mix ID) hue with lime green thin stalks. Leaflets are longer and ture. Flower onset was initiated with 12/12 day/night when narrower than most of drug cannabis varieties. plant reached approximately 16" in vegetative height. 0202 BRO5 displays vigorous hybrid character. 0218. Potential Uses of this Line. 0203 There is little or no purple on this plant until the final 0219. Potential uses of BRO5 includebut are not limited to weeks of flowering. Leaves turn deep purple with flowers medical applications, extractions, commercial raw material silvering up as time goes on. Stalks radiate a hazy or musty (chemical), fiber and pulp. urine scent. Canopy extremely sparse and topping near flow 0220 Patient Testimonials/Comments and Visual Obser ering is encouraged for even growth. Vations. 0204 Onset of Flowering and Inflorescences. 0221 Patients rave about the great experience of using 0205 Leaves are 9 and 7 leaflet patterns with 7 being BRO5. The effects are mind stimulating with some visual predominant and overall decreasing to 1 and to none in the crispness. The patients often comment that this variety is presence of female flowers. In particular, flower onset is very good for the new user because of its lower THC concentra slow with this variety. Hairy flowers are not very dense. tion and the clarity of the experience. Female flowers are spread apart due to the large internode 0222 Chemical Analysis. spacing. 0223 Low MW Terpene Analysis (136). Sesquiterpene 0206. Upon flower set, buds and supporting structures Analysis (204). Cannabinoid Analysis (>284). (stems, leaves, etc.) take longer than most to become covered with trichome bodies. Everything about this plant takes WHI2 longer. As inflorescences mature, they become more hard 0224 Description of Breeding Stock. ened and dense. In particular, the oily character of these 0225 Inflorescences were obtained for a proprietary flowers was the driving force for selection. breeding program and seeds from these inflorescences were 0207. Description of Finished Flower. isolated and put into conditions proper for their germination. 0208 BRO5 defines heady, hazy medicine with highly The seeds which germinated grew identically. The resulting functional mental effects. This variety has the structure and plants were then crossed with GO13 plants and seeds were scent of the BRO5 lines famous around the world. With aro planted and germinated for selection based on oil content of mas of spice and anise, the hashish flavor when Smoked is the plants. Plants with higher oil content were selected to enlightening. create WHI2. 0209 BRO5 is noted for mood elevation, inspiration and 0226 Hypothesized Genetics. creativity and is also likely to improve home hygiene. 0227 “Cannabis indica ssp. afghanica WLD “SB 0210 Chemotype Description for Patient. Purple'xC. indica ssp. indica NLDxC. indica ssp. kafiris 0211 Relative potency: strong. Headspace Terpenes: tanica NLDA’ pinenes, limonene. Caryophyllene content: high. 0228 Propagation and Vegetative Growth. 0212 Description of Planting, Harvesting and Processing 0229 Cuttings from WHI2 are marked by 5-finger leaflet of the Plants. sets with internode buds asymmetrically located on alternate 0213 This variety is asexually propagated via taking cut sides on main shoot. In particular, the internode space of this tings of shoots and putting them in rock wool cubes. These variety tends to be longer and stalks thinner (~4-8" veg, cubes were presoaked with pH adjusted water and kept warm decreasing flower onset). Plants are tall, stretchy and produc (-80°C.). Full trays were covered, left under 18 hours of light tive. Roots of the cuttings appear within 10-14 days. and allowed to root (7-14 days). 0230. The WHI2 grows tall and stretchy and exhibits little 0214. Upon root onset, the plantlets were transplanted into or no apical dominance. rigid 1 gallon containers filled with a proprietary soil mix A 0231 WHI2 grows with asymmetrical bud and leaf sets. and remain in 18 hours of daylight for another 14-21 days. Vegetative growth is marked by a lavish green (Munsell ID) Once root bound, plants are transplanted into rigid 3 gallon hue with green undersides and hard wood like stalks. When containers filled with proprietary soil mix B. Immediately, the healthy, fan leaves are extremely jagged and Serrations are light cycle is altered to 12/12 and flower initiating begins. The very pronounced. plants remain in 12/12 lighting until harvesting. They 0232. The stems are strong and fibrous, but extremely thin. undergo a propriety nutrient regimen and grow as undisturbed The standout quality of WHI2 is the amount of trichomes and as possible for 60-70 days depending on chemotype analysis. their density. The flower sets look frosty before most other 0215 All sun leaves are removed and plant dismantled to varieties. result in approximately 12" branches covered in inflores 0233 Stalks are vanilla spice scent. cences and trichomes. The goal in harvesting is to realize that 0234 Canopy is extremely sparse with clustered bud for we are actually harvesting trichome heads but not buds. mation. Topping extremely encouraged. Thus, great care is taken not to disturb the trichome heads and 0235. Onset of Flowering and Inflorescences. as much of the plant remains intact as possible to promote 0236 Leaves are 5 leaflet patterns with 5 being predomi even and slow drying. Slow drying followed by a one to two nant and overall decreasing to 1 and to none in the presence of month curing process. female flowers. In particular, flower onset is fast by compari 0216. Yield Data. son to most varieties. 0217. Yield determined on a per plant basis and deter 0237 Trichome density and smell are almost immediate. mined by specified cultivation techniques employed. In this Female flowers are spear-shaped, dense and thick although US 2014/029851 1 A1 Oct. 2, 2014 26 relatively large internode lengths. Again, this variety tends to 0253 Patient Testimonials/Comments and Visual Obser be more densely covered with trichome bodies than most Vations. other varieties. 0254 Very interesting from an organoleptic standpoint 0238. The flowers are compact and well-formed in the (Sweet Amsterdam flavor) and a caryophyllene content stand shape of Small pinecones. As inflorescences mature, the den point. WHI2 produces a happy laughing high, with the classic sity compact sets compound to form bright orange and silver combusted aroma of 1990’s landrace varieties of the same flowers that give way to yellow and purple Sun leaves. cannabis class. 0239 Plants are marked by unusually high oil mass con tent and extremely dense Small resinous buds. (0255 Chemical Analysis. 0240 Apical inflorescences are often smaller than lowers. 0256 Low MW Terpene Analysis (136). Sesquiterpene Inflorescences particularly are resistant to fungal infestation Analysis (204). Cannabinoid Analysis (>284). Chemical due to compact oil flowers. analysis for WHI2 plants is summarized in Tables 7 and 8. 0241. Description of Finished Flower. 0242 WHI2 (a.k.a., internally known as Heiress or “Oily TABLE 7 Heiress) was bred from a dream team of medical cannabis genetics: Northern LightsxHaze, Santa Barbara Purps, a Mid Chemical analysis of flower samples for WHI2 for Sample # 2785-6. west G-13 and the aforementioned GO13. The chemotype of THC CBD CBN this variety is indicative of this diverse genetic heritage. The aroma consists of vanilla, grapefruit, and even has petroleum Average Maximum THCA Average Maximum CBDA Maximum notes, but a rich creamy Vanilla flavor emerges when Smoked. Total content for unheated flower sample, 9% by wt. Noted for its rare combination of clarity and profound potency, it delivers functional and long lasting inspiration and O.45 ND 18.97 <0.1 ND <0.1 <0.1 positivity. Total content for unheated flower sample, mg/g 0243 Chemotype Description for Patient. 4.5 ND 189.7 <0.1 ND 0.7 O.1 0244 Relative potency: very strong. Headspace Terpenes: Estimated total content for ideally heated flower sample, % by wt. pinenes, myrcene, limonene, ocimene, linalool, terpineol. Caryophyllene content: Very high ND 12.1 ND ND <0.1 ND <0.1 0245. Description of Planting, Harvesting and Processing ND = Not determined. Note: of the Plants. Most of the acid cannabinoids (e.g., THCA) readily converts to their neutrals (THC) upon 0246 This variety is asexually propagated via taking cut heating, giving different properties, tings of shoots and putting them in rock wool cubes. These cubes were presoaked with pH adjusted water and kept warm (-80°C.). Full trays were covered, left under 18 hours of light TABLE 8 and allowed to root (7-14 days). The cannabinoid and terpene profiles, unheated, % by wt for 0247. Upon root onset, the plantlets were transplanted into Sample #: 2785-6 rigid 1 gallon containers filled with a proprietary soil mix A Cannabinoid Profile Terpenoid Profile and remain in 18 hours of daylight for another 14-21 days. % by wt % by wt Once root bound, plants are transplanted into rigid 3 gallon THCA 1897 Terpinolene O.O2 containers filled with proprietary soil mix B. Immediately, the A-THC O45 Myrcene <0.01 light cycle is altered to 12/12 and flower initiating begins. The A8-THC <0.03 Limonene <0.01 plants remain in 12/12 lighting until harvesting. They THCA-C4 O.04 Linalool O.30 undergo a propriety nutrient regimen and grow as undisturbed THCVA O.08 Caryophyllene <0.01 Oxide as possible for 60-70 days depending on chemotype analysis. THCV <0.03 Beta- 0.44 0248 All sun leaves are removed and plant dismantled to Caryophyllene result in approximately 12" branches covered in inflores CBDA O.O7 Phytol <0.01 CBD <0.03 alpha-pinene O16 cences and trichomes. The goal in harvesting is to realize that CBCA O.O7 one is actually harvesting trichome heads but not buds. CBC <0.03 Thus, great care is taken not to disturb the trichome heads and CBGA O.39 as much of the plant remains intact as possible to promote CBG O.21 CBLA <0.03 even and slow drying. Slow drying followed by a one to two CBNA <0.03 month curing process. CBN <0.03 0249. Yield Data. 0250 Yield was determined on a per plant basis and determined by specified cultivation techniques employed. In this case, indoor CEA technique following the protocol SIL4 described elsewhere herein. Flower onset was initiated with 12/12 day/night at approximately 16" in vegetative height. (0257 Description of Breeding Stock. Total biomass ~120g, finished flowers ~40 g, and/or ~30g of 0258 Inflorescences were obtained for a landrace of SIL4 seed per plant. and seeds from these inflorescences were isolated and put into 0251 Potential Uses of this Line. conditions proper for their germination. The seeds which 0252 Potential uses of WHI2 includebut are not limited to germinated grew fairly similarly. However, upon flower medical applications, extractions, commercial raw material onset, the seedlings were selected for trichome density, leaflet (e.g., chemical), fiber and pulp. width and root vigor to create SIL4. US 2014/029851 1 A1 Oct. 2, 2014 27
0259 Hypothesized Genetics. 0271 Chemotype Description for Patient. 0272 Relative potency: mild. Headspace Terpenes: 0260 “Cannabis indica ssp. afghanica WLD’ pinenes, myrcene, limonene, linalool. Caryophyllene con 0261 Propagation and Vegetative Growth. tent: medium 0262 Cuttings from SIL4 are marked by 7-finger leaflet 0273. Description of Planting, Harvesting and Processing sets with internode buds asymmetrically located on alternate of the Plants. sides on main shoot. In particular, the internode space of this 0274 This variety is asexually propagated via taking cut variety tends to be greater than that of other Silver varieties tings of shoots and putting them in rock wool cubes. These and stems harden more slowly. In particular, the cutting roots cubes were presoaked with pH adjusted water and kept warm (-80°C.). Full trays were covered, left under 18 hours of light more rapidly than other Silver varieties. In fact, the root and allowed to root (7-14 days). bodies of the plant are the most robust and vigorous of all 0275 Upon root onset, the plantlets were transplanted into cannabis plants tested in our laboratory. Root time varies with rigid 1 gallon containers filled with a proprietary soil mix A nodes appearing within 7-10 days and roots within 10-14 and remain in 18 hours of daylight for another 14-21 days. days. Once root bound, plants are transplanted into rigid 3 gallon 0263. The SIL4 grows medium in stature with stocky containers filled with proprietary soil mix B. Immediately, the branches and stalks. Even growth throughout with asym light cycle is altered to 12/12 and flower initiating begins. The metrical bud and leaf sets. Vegetative growth is marked by a plants remain in 12/12 lighting until harvesting. They deep blue-green (Munsell ID) hue with lime green thin stalks. undergo a propriety nutrient regimen and grow as undisturbed Leaflets are fat and exhibit classic recreational indica look. as possible for 60-70 days depending on chemotype analysis. These broad leaflets are indicative of this variety. Petioles are 0276 All sun leaves are removed and plant dismantled to marked by purple pointillism increasing on sides exposed to result in approximately 12" branches covered in inflores light and the end closest to palm of the leaflet set. Root bodies cences and trichomes. The goal in harvesting is to realize that are typically full and bright white. Stalks radiate a pungent we are actually harvesting trichome heads but not the buds. Smell of bubble gum coffee and green class. Canopy Thus, great care is taken not to disturb the trichome heads and extremely sparse and apical dominance can be clearly as much of the plant remains intact as possible to promote observed and removal of apical meristem often results in even and slow drying. Slow drying followed by a one to two stunted growth. Main stems may also exhibit purpling, and month curing process. inflorescences sets are large, but spread out. (0277. Yield Data. 0278 Yield was determined on a per plant basis and 0264 Onset of Flowering and Inflorescences. determined by specified cultivation techniques employed. In 0265 Leaves are 7 and 5 leaflet patterns with 3 being this case, indoor CEA technique following the protocol predominant and overall decreasing to 1 and to none in the described elsewhere herein. presence of female flowers. Female flowers are spread out due 0279 Flower onset was initiated with 12/12 day/night at to the large internode spacing. approximately 12" in vegetative height. Total biomass ~120 g, finished flowers ~30 g, and/or ~15 g of seed per plant. 0266. Upon flower set, buds and supporting structures 0280 Potential Uses of this Line. (stems, leaves, etc.) are quickly covered with an extremely 0281 Potential uses of SIL4 include but are not limited to dense field of trichome bodies. Again, this variety tends to be medical applications, extractions, commercial raw material more densely covered with trichome bodies than its parent (e.g., chemical), fiber and pulp. and other Silver varieties. In fact, the inflorescences are very 0282 Patient Testimonials/Comments and Visual Obser dense and have large calyxes covered in highly resinous vations. Patients rave about the coffee flavor and oily and glands that exhibit this variety’s distinct lemon Pine-Sol scent silver composition of SIL4. In fact, besides the mellow after only 7-10 days. effects, SIL4 is particularly noted for treating pain and inspi 0267 As inflorescences mature, the dense and compact ration. calyx clusters or flower sets give way to foxtailing and reach 0283 Chemical Analysis. ing by individual calyxes, resulting in an overall increase in 0284 Low MW Terpene Analysis (136). Sesquiterpene Surface area dedicated to trichome production. In particular, Analysis (204). Cannabinoid Analysis (>284). Chemical the oily character of these flowers set SIL4 apart from its analysis for SIL4 plants is summarized in Tables 9 and 10. parent and other Silver varieties. Textures are extremely sticky and fibrous. Stems do not break they tear, but remain TABLE 9 attached via intense fiber strands. Chemical analysis of flower samples for SIL4 for Sample # 2651-3: 0268. Description of Finished Flower. 0269. SIL4 has descended from the great Afghan hashish THC CBD CBN cannabis cultivars and is a nearly perfect choice for vapor Average Maximum THCA Average Maximum CBDA Maximum ization. The resin content delivers a range of tastes and effects with each draw. Total content for unheated flower sample, 9% by wt. 0270. The aroma consists of coffee, spice and exotic O.35 ND 1742 <0.1 ND <0.1 <0.1 incense. This variety is noted for its ability to mellow without Total content for unheated flower sample, mg/g sedation or fatigue, excellent analgesic effects and deep intro 3.5 ND 1742 <0.1 ND O.8 O.1 spection. US 2014/029851 1 A1 Oct. 2, 2014 28
TABLE 9-continued hybrid character. BLU8's stand-out quality feature is the high amount of trichomes and the high amount of oil. Stalks have Chemical analysis of flower samples for SIL4 for Sample # 2651-3: a pungent medical scent. 0295 Plant canopy is dense with large cola formation. THC CBD CBN Topping encouraged. Average Maximum THCA Average Maximum CBDA Maximum 0296. Onset of Flowering and Inflorescences. 0297 Leaves are 7 leaflet patterns with 7 being predomi Estimated total content for ideally heated flower sample, % by wt. nant and overall decreasing to 1 and to none in the presence of ND 11.0 ND ND <0.1 ND <0.1 female flowers. In particular, flower onset is medium-fast by comparison to most varieties. Trichome density and Smell are ND = Not determined. almost immediate. Female flowers are clustered to do Note: Most of the acid cannabinoids (e.g., THCA) readily converts to their neutrals (THC) upon decreased internode spacing. Again, this variety tends to be heating, giving different properties, more densely covered with trichome bodies than its parents and other varieties. In fact, the inflorescences are very dense and have large calyxes covered in highly resinous glands that TABLE 10 exhibit this variety’s distinct blueberry pine medicine/me The cannabinoid and terpene profiles, unheated, % by wt for dicinal scent after only 7-10 days. Sample #: 2651-3 0298 As inflorescences mature, the density compact sets Cannabinoid Profile Terpenoid Profile compound to form bright green and extremely oily buds. In % by wt % by wt particular, the oily character of these flowers set this its parent and phenotypes. THCA 17.42 terpinolene O.O7 A9-THC O.35 myrcene <0.01 0299. Description of Planting, Harvesting and Processing A8-THC <0.03 Limonene <0.01 of the Plants. THCA-C4 O.04 Linalool O.19 0300. This variety is asexually propagated via taking cut THCVA O.09 Caryophyllene <0.01 tings of shoots and putting them in rock wool cubes. These Oxide THCV <0.03 Beta- O.25 cubes were presoaked with pH adjusted water and kept warm Caryophyllene (-80°C.). Full trays were covered, left under 18 hours of light CBDA O.O8 Phytol <0.01 and allowed to root (7-14 days). CBD <0.03 alpha-pinene O.O1 0301 Upon root onset, the plantlets were transplanted into CBCA O.O6 CBC <0.03 rigid 1 gallon containers filled with a proprietary soil mix A CBGA O.31 and remain in 18 hours of daylight for another 14-21 days. CBG O.O6 Once root bound, plants are transplanted into rigid 3 gallon CBLA <0.07 containers filled with proprietary soil mix B. Immediately, the CBNA <0.03 light cycle is altered to 12/12 and flower initiating begins. CBN <0.03 0302) The plants remain in 12/12 lighting until harvesting. They undergo a propriety nutrient regimen and grow as undis turbed as possible for 60-70 days depending on chemotype BLU8 analysis. 0303 All sun leaves are removed and plant dismantled to 0285) Description of Breeding Stock. result in approximately 12" branches covered in inflores 0286 Inflorescences were obtained from a DJ Shorts Flo cences and trichomes. The goal in harvesting is to realize that (a.k.a. DJs Flo) pollinated by a hermaphroditic Hawaiian we are actually harvesting trichome heads but not buds. plant and seeds from these inflorescences were isolated and Thus, great care is taken not to disturb the trichome heads and put into conditions proper for their germination. as much of the plant remains intact as possible to promote 0287. The seeds which germinated grew very uniformly in even and slow drying. Slow drying followed by a one to two appearance. However, the seedlings were selected for vigor month curing process. ous phenotype with highest trichome density and oily feel of 0304 Potential Uses of this Line. resin glands to create BLU8. 0305 Potential uses of BLU8 includebut are not limited to 0288 Hypothesized Genetics. medical applications, extractions, commercial raw material 0289) “1995 Hawaiian Bag Seed:XThai'. (e.g., chemical), fiber and pulp. 0290 Propagation and Vegetative Growth. 0306 Patient Testimonials/Comments and Visual Obser 0291 Cuttings from BLU8 are marked by 7-finger leaflet Vations. sets with internode buds asymmetrically located on alternate 0307 BLU8 is very interesting from an organoleptic sides on main shoot. In particular, the internode space of this standpoint and it is unique in almost all visual categories. variety tends to be medium-stretchy (-4" veg, decreasing 0308 Chemical Analysis. flower onset). Plants are tall, robust and lanky. Cuttings root (0309 Low MW Terpene Analysis (136). Sesquiterpene within 10-14 days. Analysis (204). Cannabinoid Analysis (>284). 0292. The BLU8 grows tall and stout with mixed apical dominance. RED2 0293 BLU8 grows with asymmetrical bud and leaf sets. 0310. Description of Breeding Stock. Vegetative growth is marked by a deeper off green (Munsell 0311 Inflorescences were obtained from a random misla ID) hue with deep purple strong hollow stalks. beled dispensary clone, and selfed seeds from these plants 0294. When healthy, sun leaves are gigantic with magenta were germinated. The seeds which germinated grew very and purple under side coloring. Plants have Super vigor and similarly. The resulting seedlings were selected for vigor. US 2014/029851 1 A1 Oct. 2, 2014 29
0312 Hypothesized Genetics. light cycle is altered to 12/12 and flower initiating begins. The 0313 “Cannabis indica ssp. afghanica WLD “Cherry plants remain in 12/12 lighting until harvesting. They Afghan'xC. indica ssp. indica NLD hybrid undergo a propriety nutrient regimen and grow as undisturbed 0314 Propagation and Vegetative Growth. as possible for 60-70 days depending on chemotype analysis. 0315 Cuttings from RED2 are marked by 7-finger leaflet 0331 All sun leaves are removed and plant dismantled to sets with internode buds asymmetrically located on alternate result in approximately 12" branches covered in inflores sides on main shoot. In particular, the internode space of this cences and trichomes. The goal in harvesting is to realize that variety tends to be medium-stretchy (~4" veg, decreasing we are actually harvesting trichome heads but not buds. flower onset). The plants are tall, robust and lanky. Cuttings Thus, great care is taken not to disturb the trichome heads and root within 10-14 days. as much of the plant remains intact as possible to promote 0316 The RED2 grows tall and strong with little apical even and slow drying. Slow drying followed by a one to two dominance. RED2 grows with asymmetrical bud and leaf month curing process. SetS. 0332 Yield Data. 0317 Vegetative growth is marked by a lighter shade of 0333 Yield was determined on a per plant basis and green (Munsell ID) hue with deep purple strong hollow determined by specified cultivation techniques employed. In stalks. When healthy, sun leaves are point upward toward this case, indoor CEA technique following the protocol light source. described elsewhere herein. Flower onset was initiated with 0318. The stems are strong and fibrous. The plants are 12/12 day/night at approximately 16" in vegetative height. Super vigorous and hybrid in character. The stand-out quality Total biomass ~180 g, finished flowers ~60 g, and/or ~50 g of is the high amount of trichomes and the high amount of oil. seed per plant. Stalks have a Sweet scent. Canopy is dense with large cola 0334 Potential Uses of this Line. formation. Topping encouraged. 0335 Potential uses of RED2 includebut are not limited to 0319. Onset of Flowering and Inflorescences. medical applications, extractions, commercial raw material 0320 Leaves are 7 leaflet patterns with 7 being predomi (e.g., chemical), fiber and pulp. nant and overall decreasing to 1 and to none in the presence of 0336 Patient Testimonials/Comments and Visual Obser female flowers. In particular, flower onset is medium-fast by Vations. comparison to most varieties. 0337. Noted as being very interesting from an organolep 0321 Trichome density and smell are almost immediate. tic standpoint. RED2 is unique in almost all visual categories. Female flowers are clustered to do decreased internode spac 0338 Chemical Analysis. ing. Again, this variety tends to be more densely covered with 0339 Low MW Terpene Analysis (136). Sesquiterpene trichome bodies than its parents and other varieties. In fact, Analysis (204). Cannabinoid Analysis (>284). Chemical the inflorescences are very dense and have large calyxes analysis for RED2 plants is summarized in Tables 11 and 12. covered in highly resinous glands that exhibit this variety's distinct blueberry pine medicine/medicinal scent after only TABLE 11 7-10 days. 0322. As inflorescences mature, the dense and compact Chemical analysis of flower Samples for RED2 for Sample # 2785-4: calyx clusters or flower sets form bright green and extremely THC CBD CBN oily buds. In particular, the oily character of these flowers set this variety apart from its parent and phenotypes. Average Maximum THCA Average Maximum CBDA Maximum 0323 Description of Finished Flower. Total content for unheated flower sample, 9% by wt. 0324 RED2 combines a beautifully sweet cherry WLD Afghan with a NLD to deliver a strong, cheerful, dreamy O.26 ND 17.10 <0.1 ND <0.1 <0.1 psychoactivity. This variety produces a pleasant silliness and Total content for unheated flower sample, mg/g a where'd I put my keys memory effect and obliterates most 2.6 ND 1710 <0.1 ND 0.7 O.1 patient troubles. Estimated total content for ideally heated flower sample, % by wt. 0325 Aroma consists of cherry cough drops, fresh straw berries and just a hint of spice. RED2 is often noted for ND 10.8 ND ND <0.1 ND <0.1 long-lasting effects and positive mood impact. ND = Not determined. 0326 Chemotype Description for Patient. Note: Most of the acid cannabinoids (e.g., THCA) readily converts to their neutrals (THC) upon 0327 Relative potency: strong. Headspace Terpenes: heating, giving different properties, pinenes, myrcene, limonene, and linalool. Caryophyllene content: high 0328. Description of Planting, Harvesting and Processing TABLE 12 of the Plants. The cannabinoid profiles, unheated, % by wt for Sample #: 2785-4 0329. This variety is asexually propagated via taking cut Cannabinoid Profile % by wit tings of shoots and putting them in rock wool cubes. These THCA 17.10 cubes were presoaked with pH adjusted water and kept warm A9-THC O.26 (-80°C.). Full trays were covered, left under 18 hours of light A8-THC <0.03 and allowed to root (7-14 days). THCA-C4 O.O3 0330 Upon root onset, the plantlets were transplanted into THCVA O.21 THCV <0.03 rigid 1 gallon containers filled with a proprietary soil mix A CBDA O.O7 and remain in 18 hours of daylight for another 14-21 days. CBD <0.03 Once root bound, plants are transplanted into rigid 3 gallon CBCA O.O6 containers filled with proprietary soil mix B. Immediately, the US 2014/029851 1 A1 Oct. 2, 2014 30
TABLE 12-continued 0354) Description of Finished Flower. 0355 GRE1 defines sweet, delicious medicine/medicinal The cannabinoid profiles, unheated, % by wt for Sample #: 2785-4 with functional mental effects. This variety has resin produc Cannabinoid Profile % by wit tion akin to Afghan and psychoactivity reminiscent of origi CBC <0.03 nal Green class. CBGA <0.03 CBG O.39 0356 GRE1 has aromas of citrus, brown sugar, and CBLA <0.03 banana nut bread combine to produce a fantastic fruity hash CBNA <0.03 ish flavor when smoked. It is noted for mood elevation and CBN <0.03 daytime bursts of energy that provide for short-term pain relief. 0357 Chemotype Description for Patient. GRE1 0358 Relative potency: Strong. Headspace Terpenes: pinenes, myrcene, limonene, and ocimene. Caryophyllene 0340 Description of Breeding Stock. content: Medium. 0341 Inflorescences were obtained and isolated and put 0359. Description of Planting, Harvesting and Processing into conditions proper for their germination. of the Plants. 0342. The seeds which germinated grew identically. How 0360 This variety is asexually propagated via taking cut ever, the seedlings were selected for the phenotype that is tings of shoots and putting them in rock wool cubes. These more densely covered in trichomes, where the oil content of cubes were presoaked with pH adjusted water and kept warm the gland heads was higher than other phenotypes of this (-80°C.). Full trays were covered, left under 18 hours of light variety. and allowed to root (7-14 days). 0343 Hypothesized Genetics. 0361 Upon root onset, the plantlets were transplanted into 0344). “Cannabis indica ssp. afghanica WLD “Afghan rigid 1 gallon containers filled with a proprietary soil mix A #1'xC. indica ssp. indica NLD hybrid and remain in 18 hours of daylight for another 14-21 days. (0345 Propagation and Vegetative Growth. Once root bound, plants are transplanted into rigid 3 gallon containers filled with proprietary soil mix B. Immediately, the 0346 Cuttings from GRE1 are marked by 9-finger very light cycle is altered to 12/12 and flower initiating begins. The thin leaflet sets with internode buds asymmetrically located plants remain in 12/12 lighting until harvesting. They on alternate sides on main shoot. In particular, the internode space of this variety tends to be extremely large. Plants are undergo a propriety nutrient regimen and grow as undisturbed tall, frail and stretchy. Cuttings root appears within 10-14 as possible for 60-70 days depending on chemotype analysis. days. 0362 All sun leaves are removed and plant dismantled to result in approximately 12" branches covered in inflores 0347 GRE1 grows tall and stretchy with flimsy stems and cences and trichomes. The goal in harvesting is to realize that embodies what it means to be a true hybrid. we are actually harvesting trichome heads but not buds. 0348 GRE1 grows with asymmetrical bud and leaf sets. Thus, great care is taken not to disturb the trichome heads and Vegetative growth is marked by a lush green (Munsell ID) hue as much of the plant remains intact as possible to promote with lime green thin stalks. even and slow drying. Slow drying followed by a one to two 0349 Leaflets are longer and thinner than varieties. Plants month curing process. have a vigorous hybrid character. GRE1 has little or no purple 0363 Yield Data. color on the plant. The stand-out quality is the high amount of 0364 Yield was determined on a per plant basis and trichomes and the high amount of oil. Plant stalks have a determined by specified cultivation techniques employed. In Sweet citrus creamsicle scent. this case, indoor CEA technique following the protocol 0350 Plant canopy is dense and even topping near flow described elsewhere herein. ering is encouraged for even growth. 0365 Flower onset was initiated with 12/12 day/night at 0351 Onset of Flowering and Inflorescences. approximately 16" in vegetative height. Total biomass ~160 0352 Leaves are 9 and 7 leaflet patterns with 7 being g, finished flowers -50 g, and/or ~50 g of seed per plant. predominant and overall decreasing to 1 and to none in the 0366 Potential Uses of this Line. presence of female flowers. In particular, flower onset is fast 0367 Potential uses of GRE1 includebut are not limited to by comparison to most varieties. medical applications, extractions, commercial raw material 0353 Trichome density and smell are almost immediate. (e.g., chemical), fiber and pulp. Female flowers are clustered to do decreased internode spac 0368 Patient Testimonials/Comments and Visual Obser ing. Again, this variety tends to be more densely covered with Vations. trichome bodies than its parent and other green class varieties. 0369 Patients rave about the great experience of using this In fact, the inflorescences are very dense and have large plant. The flowers of GRE1 consistently produce approxi calyxes covered in highly resinous glands that exhibit this variety’s distinct green class creamsicle scent after only 7-10 mately 2.0% CBGA in finished flowers. Its wonderful smell/ days. As inflorescences mature, the density compact sets taste is patients major reason for appeal. compound to form orange and bright green extremely oily 0370 Chemical Analysis. buds. In particular, the oily character of these flowers set this 0371 Low MW Terpene Analysis (136). Sesquiterpene green class apart from its parent and other green class variet Analysis (204). Cannabinoid Analysis (>284). Chemical 1CS analysis for GRE1 plants is summarized in Tables 13 and 14. US 2014/029851 1 A1 Oct. 2, 2014 31
TABLE 13 Vegetative growth is marked by a deeper off green (Munsell ID) hue with lime green thin stalks. Leaflets are longer and Chemical analysis of flower samples for GRE1 for Sample # 2651-4: thinner than varieties. When healthy, Sun leaves are gigantic. It has vigorous hybrid character. The stand-out quality is the THC CBD CBN high amount of trichomes and the high amount of oil. There is Average Maximum THCA Average Maximum CBDA Maximum an extremely high cannabinoid content in PUR3. Stalks have a Sweet dank scent. Canopy dense and do not need to top. Total content for unheated flower sample, 9% by wt. 0380. Onset of Flowering and Inflorescences. O.74 ND 2O.S2 <0.1 ND <0.1 <0.1 0381 Leaves are 7 leaflet patterns with 7 being predomi Total content for unheated flower sample, mg/g nant and overall decreasing to 1 and to none in the presence of 7.4 ND 205.2 <0.1 ND O.9 O.2 female flowers. In particular, flower onset is fast by compari Estimated total content for ideally heated flower sample, % by wt. son to most varieties. 0382 Trichome density and smell are almost immediate. ND 13.3 ND ND <0.1 ND <0.1 Female flowers are clustered to do decreased internode spac ND = Not determined. ing. Again, this variety tends to be more densely covered with Note: trichome bodies than its parents and other purple varieties. In Most of the acid cannabinoids (e.g., THCA) readily converts to their neutrals (THC) upon fact, the inflorescences are very dense and have large calyxes heating, giving different properties, covered in highly resinous glands that exhibit this variety's distinct grape lollipop scent after only 7-10 days. As inflores TABLE 1.4 cences mature, the density compact sets compound to form deep purple and darkgreen extremely oily buds. In particular, The cannabinoid and terpene profiles, unheated, % by wt for the oily character of these flowers set this purple apart from its Sample #: 2651-4 parent and other green class varieties. Cannabinoid Profile Terpenoid Profile (0383. Description of Finished Flower. % by wt % by wt 0384 PUR3 defines sweet, delicious grape flavored medi THCA 2O.S2 terpinolene <0.01 cine with functional mental effects and pain relief. This vari A9-THC O.74 myrcene <0.01 ety has resin production akin to Afghan and psychoactivity A8-THC <0.03 Limonene <0.01 reminiscent of the PUR3. PUR3 has aromas of grape, Sweet THCA-C4 O.04 Linalool O.12 THCVA O.18 Caryophylene O.O2 Sugar, and dank which all combine to produce a fantastic Oxide grape flavor when Smoked. It is noted for mood elevation, THCV <0.03 Beta- O.22 short-term pain relief and hunger stimulation. Caryophyllene (0385 Chemotype Description for Patient. CBDA O.09 Phytol <0.01 CBD <0.03 alpha-pinene O.17 0386 Relative potency: very strong. Headspace Terpenes: CBCA O16 pinenes, myrcene, and limonene. Caryophyllene content: CBC <0.03 high. CBGA 148 0387 Description of Planting, Harvesting and Processing CBG O.28 CBLA O.15 of the Plants. CBNA <0.03 0388. This variety is asexually propagated via taking cut CBN <0.03 tings of shoots and putting them in rock wool cubes. These cubes were presoaked with pH adjusted water and kept warm (-80°C.). Full trays were covered, left under 18 hours of light PUR3 and allowed to root (7-14 days). 0389. Upon root onset, the plantlets were transplanted into 0372. Description of Breeding Stock. rigid 1 gallon containers filled with a proprietary soil mix A 0373) Inflorescences were obtained for a landrace of and remain in 18 hours of daylight for another 14-21 days. purple class pollinated with a hermaphroditic purple class Once root bound, plants are transplanted into rigid 3 gallon variety and seeds from these inflorescences were isolated and containers filled with proprietary soil mix B. Immediately, the put into conditions proper for their germination. light cycle is altered to 12/12 and flower initiating begins. The 0374. The seeds which germinated grew very uniformly in plants remain in 12/12 lighting until harvesting. They appearance. However, upon flower onset, the seedlings were undergo a propriety nutrient regimen and grow as undisturbed selected for the two phenotypes that most smelled like grape as possible for 60-70 days depending on chemotype analysis. and dank, and producing flowers with the highest trichome 0390 All sun leaves are removed and plant dismantled to density and robust examples of these two phenotypes were result in approximately 12" branches covered in inflores subsequently crossed to create PUR3. cences and trichomes. The goal in harvesting is to realize that 0375 Hypothesized Genetics. we are actually harvesting trichome heads but not buds. 0376 “2007 SB PUP1x2009 PPS7. Thus, great care is taken not to disturb the trichome heads and 0377 Propagation and Vegetative Growth. as much of the plant remains intact as possible to promote 0378 Cuttings from PUR3 are marked by 7-finger leaflet even and slow drying. Slow drying followed by a one to two sets with internode buds asymmetrically located on alternate month curing process. sides on main shoot. In particular, the internode space of this 0391. Yield Data. variety tends to be decreased. Short, squatty and bushy. Cut 0392 Yield was determined on a per plant basis and tings root within 10-14 days. determined by specified cultivation techniques employed. In 0379 The PUR3 grows stout in the traditional Christmas this case, indoor CEA technique following the protocol tree' shape. PUR3 grows with asymmetrical bud and leaf sets. described elsewhere herein. US 2014/029851 1 A1 Oct. 2, 2014 32
0393 Flower onset was initiated with 12/12 day/night at for countering the debilitating aspects of many medical con approximately 16" in vegetative height. Total biomass ~160 ditions. A complex aroma of spicy spruce and lemon peel g, finished flowers -50 g, and/or -50 g of seed per plant. release a cornucopia of Sweet and spicy piney flavors when 0394 Potential Uses of this Line. Smoked. It is often characterized by a clear head, accompa 0395 Potential uses of PUR3 include but are not limited to nied by mood elevation. medical applications, extractions, commercial raw material 0412 Chemotype Description for Patient. (e.g., chemical), fiber and pulp. 0413 Relative potency: very strong. Headspace Terpenes: 0396 Patient Testimonials/Comments and Visual Obser pinenes, myrcene, limonene, ocimene, and terpinolene. Vations. Caryophyllene content: medium. 0397 Patients rave about the great experience. This flower 0414. Description of Planting, Harvesting and Processing consistently produces approximately 2.0% CBGA in finished of the Plants. flowers. Its wonderful smell/taste is patients major reason for 0415. This variety is asexually propagated via taking cut appeal. tings of shoots and putting them in rock wool cubes. These 0398 Chemical Analysis. cubes were presoaked with pH adjusted water and kept warm 0399. Low MW Terpene Analysis (136). Sesquiterpene (-80°C.). Full trays were covered, left under 18 hours of light Analysis (204). Cannabinoid Analysis (>284). and allowed to root (7-14 days). Upon root onset, the plantlets were transplanted into rigid 1 gallon containers filled with a YEL3 proprietary soil mix A and remain in 18 hours of daylight for 0400. Description of Breeding Stock. another 14-21 days. Once root bound, plants are transplanted 04.01 Inflorescences were obtained from an unknown lan into rigid 3 gallon containers filled with proprietary soil mix drace. Seeds from these inflorescences were isolated and put B. Immediately, the light cycle is altered to 12/12 and flower into conditions proper for their germination. The seeds which initiating begins. The plants remain in 12/12 lighting until germinated grew uniformly in appearance. However, the harvesting. They undergo a propriety nutrient regimen and seedlings were selected for their narrow-leafleted tropical grow as undisturbed as possible for 60-70 days depending on cannabis morphology and pinene production to create YEL3. chemotype analysis. 0402 Hypothesized Genetics. 0416 All sun leaves are removed and plant dismantled to 0403. “Cannabis indica ssp. indica NLD “Thai'xC. result in approximately 12" branches covered in inflores indica ssp. indica NLD “Highland Mexican.” cences and trichomes. The goal in harvesting is to realize that 0404 Propagation and Vegetative Growth. we are actually harvesting trichome heads but not buds. 04.05 Cuttings from YEL3 are marked by 9-finger leaflet Thus, great care is taken not to disturb the trichome heads and sets with internode buds asymmetrically located on alternate as much of the plant remains intact as possible to promote sides on main shoot. In particular, the internode space of this even and slow drying. Slow drying followed by a one to two variety tends to be lengthy and stretchy (-4" Veg, decreasing month curing process. flower onset). The plants are tall, robust and lanky. Cuttings 0417. Yield Data. root within 10-14 days. 0418 Yield was determined on a perplant basis using the 0406. The YEL3 grows tall and strong with pronounced specified cultivation techniques employed. In this case, apical dominance. YEL3 grows with asymmetrical bud and indoor CEA technique following the protocol described else leaf sets. Vegetative growth is marked by a lighter dark green where herein. Flower onset was initiated with 12/12 day/night (Munsell ID) hue with purple undersides and strong hollow at approximately 16" in vegetative height. Total biomass stalks. When healthy, sun leaves are point upward toward ~120g, finished flowers ~40 g, and/or -30g of seed perplant. light source at twisted angles. The stems are strong and fibrous. The stand-out quality is the high amount of trichomes 0419 Potential Uses of this Line. and the high amount of oil. YEL3 has stalks with a sweet 0420 Potential uses of YEL3 include but are not limited to scent. Plant canopy is sparse with scattered bud formation. medical applications, extractions, commercial raw material Topping encouraged. (e.g., chemical), fiber and pulp. 0407 Onset of Flowering and Inflorescences. 0421. Patient Testimonials/Comments and Visual Obser 0408 Leaves are 9 leaflet patterns with 9 being predomi Vations. nant and overall decreasing to 1 and to none in the presence of 0422 Plants have a very interesting from an organoleptic female flowers. In particular, flower onset is fast by compari standpoint and are unique in almost all visual categories. son to most varieties. 0423 Chemical Analysis. 04.09 Trichome density and smell are almost immediate. 0424 Low MW Terpene Analysis (136). Sesquiterpene Female flowers are sparse due to large internode spacing. Analysis (204). Cannabinoid Analysis (>284). Chemical Again, this variety tends to be more densely covered with analysis for YEL3 plants is summarized in Tables 15-17. trichome bodies than its parents and other varieties. The flow ers are not compact or well-formed. Inflorescences are spirals TABLE 1.5 of individual foxtails that form a coral looking structure. Although buds are made of individual spirals, the inflores Chemical analysis of flower samples for YEL3 for Sample # 2788-1: cences are dense and tightly packed. As inflorescences mature, the density compact sets compound to form bright THC CBD CBN green and extremely oily buds. Average Maximum THCA Average Maximum CBDA Maximum 0410. Description of Finished Flower. 0411 YEL3 has descended from the great Oaxacan and Total content for unheated flower sample, 9% by wt. That cannabis landrace plants of the 1970s. This variety O.17 ND 18.51 <0.1 ND O16 <0.1 delivers an intense “up' stimulating effect that can be great US 2014/029851 1 A1 Oct. 2, 2014 33
TABLE 15-continued germinated grew uniformly in appearance. However, the seedlings were selected for trichome density and hybrid leaf Chemical analysis of flower samples for YEL3 for Sample # 2788-1: morphology to create WHI3. Hypothesized Genetics. “Can nabis indica ssp. afghanica WLD Afghan #1'xCannabis THC CBD CBN indica ssp. indica NLD “Brazilian'xC. indica ssp. indica Average Maximum THCA Average Maximum CBDA Maximum NLD “Indian. 0427 Propagation and Vegetative Growth. Total content for unheated flower sample, mg/g 0428 Cuttings from WHI3 are marked by 5-finger leaflet 1.7 ND 185.1 <0.1 ND 1.6 <0.1 sets with internode buds asymmetrically located on alternate Estimated total content for ideally heated flower sample, % by wt. sides on main shoot. In particular, the internode space of this variety tends to be shorter and stout (-2-4" Veg, decreasing ND 11.5 ND ND O.1 ND <0.1 flower onset). The plants are short, robust and bushy. Cuttings ND = Not determined. root within 10-14 days. Note: Most of the acid cannabinoids (e.g., THCA) readily converts to their neutrals (THC) upon 0429. The WHI3 grows short and bushy with classic heating, giving different properties, Christmas tree apical dominance. WHI3 grows with asym metrical bud and leaf sets. Vegetative growth is marked by a dark green (Munsell ID) hue with green undersides and hard TABLE 16 wood like stalks. When healthy, sun leaves are point upward The cannabinoid and terpene profiles, unheated, % by wt for toward light Source and reach. The stems are strong and Sample H: 2788-1 fibrous. The stand out quality is the high amount of trichomes and the high amount of oil. The flower sets look white before Cannabinoid Profile Terpenoid Profile most other varieties. Stalks are sweet scent. Plant canopy is % by wt % by wt dense with clustered bud formation. Topping discouraged. THCA 18.51 terpinolene <0.01 0430. Onset of Flowering and Inflorescences. A9-THC O.17 myrcene O16 A8-THC <0.03 Limonene <0.01 0431 Leaves are 5 leaflet patterns with 5 being predomi THCA-C4 O.O3 Linalool O.O2 nant and overall decreasing to 1 and to none in the presence of THCVA O.O7 Caryophyllene <0.01 female flowers. In particular, flower onset is fast by compari Oxide son to most varieties. THCV <0.03 Beta- O.25 Caryophylene 0432 Trichome density and smell are almost immediate. CBDA O16 Phytol <0.01 Female flowers are dense and thick due to relatively small CBD <0.03 alpha-pinene O.O6 internode lengths. Again, this variety tends to be more CBCA O.S6 densely covered with trichome bodies than other varieties in CBC <0.03 CBGA 1.13 the white class. The flowers are compact and well-formed in CBG O.82 the shape of pinecones. Pistils are fat and of high density. As CBLA <0.03 inflorescences mature, the density compact sets compound to CBNA <0.03 form bright neon-green flowers that give way to red-orange CBN <0.03 hair. It is marked by unusually high sesquiterpene content and extremely resinous buds. Inflorescences are subject to fungal infestation due to large size and extreme density. TABLE 17 0433. Description of Finished Flower. 0434 WHI3 produces prodigious amounts of psychoac Chemical analysis of flower samples for YEL3 for Sample # 2785-1: tive resin. This variety was derived from Brazilian, Indian, THC CBD CBN and Afghan gene pools. Its aroma of green classy, balsamic, pineapple gazpacho delivers a Sweet, hashy flavor when Average Maximum THCA Average Maximum CBDA Maximum Smoked. It is noted for fast-onset psychoactivity reminiscent of traditional cannabis experiences that will leave you right Total content for unheated flower sample, 9% by wt. where it found you. Happiness induced pain relief and con O.22 ND 19.23 <0.1 ND O.13 <0.1 siderable relaxation. Total content for unheated flower sample, mg/g 0435 Chemotype Description for Patient. 2.2 ND 1923 <0.1 ND 1.3 O.2 0436 Relative potency: very strong. Headspace Terpenes: Estimated total content for ideally heated flower sample, % by wt. pinenes, myrcene, limonene, humulene, and naphthalene. Caryophyllene content: Very high ND 12.0 ND ND <0.1 ND <0.1 0437. Description of Planting, Harvesting and Processing ND = Not determined. of the Plants. Note: 0438. This variety is asexually propagated via taking cut Most of the acid cannabinoids (e.g., THCA) readily converts to their neutrals (THC) upon heating, giving different properties, tings of shoots and putting them in rock wool cubes. These cubes were presoaked with pH adjusted water and kept warm (-80°C.). Full trays were covered, left under 18 hours of light WHI3 and allowed to root (7-14 days). 0439 Upon root onset, the plantlets were transplanted into 0425 Description of Breeding Stock. rigid 1 gallon containers filled with a proprietary soil mix A 0426 Inflorescences were obtained for an unknown lan and remain in 18 hours of daylight for another 14-21 days. drace. Seeds from these inflorescences were isolated and put Once root bound, plants are transplanted into rigid 3 gallon into conditions proper for their germination. The seeds which containers filled with proprietary soil mix B. Immediately, the US 2014/029851 1 A1 Oct. 2, 2014 34 light cycle is altered to 12/12 and flower initiating begins. The TABLE 19-continued plants remain in 12/12 lighting until harvesting. They undergo a propriety nutrient regimen and grow as undisturbed The cannabinoid and terpene profiles, unheated, % by wt for as possible for 60-70 days depending on chemotype analysis. Sample #: 2651-2 0440 All sun leaves are removed and plant dismantled to Cannabinoid Profile Terpenoid Profile result in approximately 12" branches covered in inflores % by wt % by wt cences and trichomes. The goal in harvesting is to realize that THCA-C4 O.04 Linalool O.O7 we are actually harvesting trichome heads but not buds. THCVA O.09 Caryophyllene <0.01 Thus, great care is taken not to disturb the trichome heads and Oxide THCV <0.03 Beta- O.15 as much of the plant remains intact as possible to promote Caryophyllene even and slow drying. Slow drying followed by a one to two CBDA O.09 Phytol <0.01 month curing process. CBD <0.03 alpha-pinene O.31 0441. Yield Data. CBCA O.08 CBC <0.03 0442. Yield was determined on a per plant basis and CBGA O.19 determined by specified cultivation techniques employed. In CBG O16 this case, indoor CEA technique following the protocol CBLA O.04 described elsewhere herein. Flower onset was initiated with CBNA <0.03 12/12 day/night at approximately 16" in vegetative height. CBN <0.03 Total biomass ~140 g, finished flowers -50 g, and/or ~50 g of seed per plant. 0443 Potential Uses of this Line. Example 2 0444 Potential uses of WHI3 include but are not limited to medical applications, extractions, commercial raw material Development of Medical Cannabis Varieties (e.g., chemical), fiber and pulp. 0449 In another objective of the present invention, the 0445 Patient Testimonials/Comments and Visual Obser Classes of Cannabis Varieties of Example 1 are used in can Vations. nabis breeding programs to develop Medical Cannabis plants 0446 Very interesting from an organoleptic standpoint and varieties. Furthermore, the Medical Cannabis varieties (Sweet Amsterdam flavor) and a caryophyllene content stand developed according to the present invention have specific point. Happy laughing high. WHI3 has the burnt scent of flavor(s) in accordance with one of the classes of Classes of 1990’s landraces. Cannabis Varieties as discussed above. 0447 Chemical Analysis. 0450. The general idea behind this approach was in 0448 Low MW Terpene Analysis (136). Sesquiterpene response to the fact that currently available cannabis varieties Analysis (204). Cannabinoid Analysis (>284). Chemical have been skewed towards THC production, which has analysis for WHI3 plants is summarized in Tables 18 and 19. increased the likelihood of adverse effects from the elevated levels of psychoactivity that these high-THC varieties pro TABLE 18 duce. 0451 Contemporary illicit “recreational marijuana culti Chemical analysis of flower samples for WHI3 for Sample # 2651-2: vars have been exclusively bred and selected primarily for their THC acid content, secondarily for their terpenoid aroma THC CBD CBN and flavor chemistry, and rarely for their production of the Average Maximum THCA Average Maximum CBDA Maximum other cannabinoid acids, such as CBDA. 0452 Cannabidiol (CBD), a cannabinoid that is rare in Total content for unheated flower sample, 9% by wt. contemporary medical cannabis varieties, has been shown to 0.27 ND 1896 <0.1 ND <0.1 <0.1 reduce and modulate the psychoactivity of THC and also Total content for unheated flower sample, mg/g reduce some of THC's other adverse effects including tachy 2.7 ND 1896 <0.1 ND O.9 O.1 cardia, anxiety, memory effects, etc. There is some evidence Estimated total content for ideally heated flower sample, % by wt. that CBD may reduce the buildup of tolerance to the effects of THC and also reduce the likelihood of cannabis dependency. ND 11.9 ND ND <0.1 ND <0.1 0453 The breeding programs of the present invention ND = Not determined. were designed to combine lower THC with higher CBD so as Note: to produce medical cannabis varieties. Furthermore, the Most of the acid cannabinoids (e.g., THCA) readily converts to their neutrals (THC) upon medical cannabis varieties of the present invention were addi heating, giving different properties, tionally selected for their ability to produce fragrant terpe noids that are appealing to patients and that may also provide TABLE 19 a pharmacological activity that modifies, enhances or ame liorates the effects of THC. In contrast, publicly-available The cannabinoid and terpene profiles, unheated, % by wt for contemporary hemp varieties that are high in CBD do not Sample #: 2651-2 produce the pleasing organoleptic attributes of contemporary Cannabinoid Profile Terpenoid Profile high-THC marijuana cultivars. Thus, an objective of the % by wt % by wt present invention is to combine lower THC with higher CBD THCA 18.96 terpinolene <0.01 So as to produce medical cannabis varieties with these pleas A-THC 0.27 myrcene <0.01 ing aromas and flavors. A8-THC <0.3 Limonene <0.01 0454 Varieties of medical cannabis with determined cannabinoid content and increased essential oil content are US 2014/029851 1 A1 Oct. 2, 2014 designed according to the present invention so as to reduce the include but are not limited to larger plant size (i.e., greater adverse effects of THC and Smoking while increasing orga bulk or biomass), higher production of flower buds, larger noleptic appeal. flowers, more trichomes, shorter plant stature, ability to tol 0455 One embodiment of the present invention is to pro erate lower and/or higher growing temperatures, greater ger duce medical cannabis varieties with high essential oil con mination percentage, greater seedling vigor, more efficient tent, in particularly, mono- and sesquiterpenes. While THCA water usage, disease resistance, pest resistance, and other is of importance, the entourage effects of these terpenoids in desirable agronomic and production traits. Foran overview of concert with proper dosage of cannabinoids area focus of the diseases and pests of importance to cannabis production see present invention. The breeding objectives of the present Clarke et al. (2000) Hemp Diseases and Pests. Management invention are opposite to the face of modern recreational and Biological Control: An Advanced Treatise (CABI Pub marijuana breeding strategies which have focused almost lishing). solely on breeding for higher levels of THCA content alone. 0459. In an alternative version of this breeding regime the 0456. According to the present invention a Class variety is selected F2 progeny are backcrossed to the Class variety as crossed to a CBD line to produce F1 seed which were grown the recurrent parent. Selection for desirable phenotypes and/ to produce F1 progeny. Five CBD lines were chosen to use in or genotypes can be conducted after this initial backcross, the initial breeding program: CBD1, CBD2, CBD3, CBD4 after any Subsequent backcross (e.g., progeny obtained after and CBD5. According to the present invention, each of these 2, 3, 4, 5, 6, 7, 8, 9 or more backcrosses). Selected lines will CBD lines is crossed to one or more cannabis varieties in a have a total THC contente2% but s90.0%, a total CBD con specific Class. So, for example, one or more GOLD Class tents 1.5%, and an aroma and flavor(s) typical of its class. In varieties are crossed to each of CBD1, CBD2, CBD3, CBD4 another version of this breeding scheme selected lines can be and CBD5 to produce five F1 populations per Class variety to chosen to have a total THC:CBD ratio of 8:1 and approaching create GOLD ClassXCBD combinations. Table 20 is a list of 1:1, and an aroma and flavor(s) typical of its class. The lines the iterations for each of the ClassXCBD crosses. can also be further selected for a specific content of certain TABLE 20
List of Class X CBD crosses.
CBD1 Crosses CBD2 Crosses CBD3 Crosses CBD4 Crosses CBDS Crosses GOLD X CBD1 GOLD X CBD2 GOLDX CBD3 GOLD X CBD4 GOLDX CBD5 BRONZEX CBD1 BRONZEX CBD2 BRONZEX CBD3 BRONZEX CBD4 BRONZEX CBDS WHITEX CBD1 WHITEX CBD2 WHITEX CBD3 WHITEX CBD4 WHITEX CBD5 FUSCLAX CBD1 FUSCLAX CBD2 FUSCLAX CBD3 FUSCLAX CBD4 FUSCLAX CBDS ORANGEX CBD1 ORANGEX CBD2 ORANGEX CBD3 ORANGEX CBD4 ORANGEX CBDS RED X CBD1 RED X CBD2 RED X CBD3 RED X CBD4 RED X CBDS YELLOW X CBD1 YELLOW X CBD2 YELLOW X CBD3 YELLOW X CBD4 YELLOW X CBDS SILVERX CBD1 SILVERX CBD2 SILVERX CBD3 SILVERX CBD4 SILVERX CBDS PURPLE X CBD1 PURPLE X CBD2 PURPLE X CBD3 PURPLE X CBD4 PURPLE X CBDS GRAY X CBD1 GRAY X CBD2 GRAY X CBD3 GRAY X CBD4 GRAYX CBDS JADEX CBD1 JADEX CBD2 JADEX CBD3 JADEX CBD4 JADEX CBDS BLUEX CBD1 BLUEX CBD2 BLUEX CBD3 BLUEX CBD4 BLUEX CBDS BLACKX CBD1 BLACKX CBD2 BLACKX CBD3 BLACKX CBD4 BLACKX CBDS GREENX CBD1 GREENX CBD2 GREENX CBD3 GREENX CBD4 GREENX CBDS MISCX CBD1 MISCX CBD2 MISCX CBD3 MISCX CBD4 MISCX CBDS
0457 In one representative version of this breeding other cannabinoids and/or of certain terpenes/terpenoids, regime the resultant F1 progeny can be selfed to produce F2 and/or for additional phenotypic and genotypic characteris seed which are grown to produce F2 progeny. Selection for tics. desirable phenotypes and/or genotypes can be conducted 0460 The progeny resulting from any selection stage of within the F2 progeny since the selections can be maintained either the selfing or backcrossing versions of the breeding (i.e., fixed) via asexual reproduction. Alternatively, the F2 regimes of the present invention can be asexually reproduced progeny can be crossed among themselves to produce a so as to fix and maintain the desirable THC content, CBD bulked F3 population from which desired progeny can be content, the aroma and flavor(s) typical of the desired class, selected and/or further generations of crossing can be con and the other desirable phenotypic and/or genotypic charac ducted. Regardless of the exact selfing/selection procedure, teristics. The resultant selected lines will be designated as selected lines can be chosen so as to have a total THC content classical medical cannabis varieties. but s90.0%, a total CBD contenta 1.5%, and an aroma and 0461 The progeny resulting from any stage of either the flavor(s) typical of its class. In another version of the present selfing or backcrossing versions of this regime can also be invention, regardless of the exact selfing/selection procedure, crossed to other cannabis plants/varieties within, between or selected lines can be chosen so as to have a total THC:CBD among the various classes of cannabis so as to produce addi ratio of 8:1 and approaching 1:1, and an aroma and flavor(s) tional plants for selection and maintenance through asexual typical of its class. reproduction. In this way, medical cannabis varieties with 0458. The lines can also be further selected for a specific various, desired flavor combinations can be produced and content of certain other cannabinoids and/or of certainterpe Subsequently maintained through asexual reproduction. nes/terpenoids, and/or for additional phenotypic and geno 0462. The resultant medical cannabis plants of the present typic characteristics. Desirable phenotypic characteristics invention also generally have more oil content per plant than US 2014/029851 1 A1 Oct. 2, 2014 36 contemporary marijuana varieties. More oil per plant means will again require self fertilization to create a segregating less plant matter is required per treatment/administration, population, and Subsequent screening of the segregating thereby also further minimizing any health risks for medical population to eliminate chemotype I and chemotype II plants, cannabis Smokers. only retaining the chemotype III plants that approach the 0463 Breeding plants with increased CBDA content will ideotype. alleviate most of the commonly recognized real and per 0470 Backcross Breeding Scheme. ceived adverse effects of high THC cannabis. A direct result 0471. The following backcross breeding procedure will of increased CBDA is lower THC content because THCA also be accomplished according to the present invention: synthase and CBDA synthase are allelic. Thus, another objec tive of the present invention is to create medical cannabis P1 (GOLD Line (GO13)xP2 (CBD Line (CBD1201) varieties with an optimal dose of THCA and resulting in the most efficacious ratio of CBDA:THCA. 0472. The hybrid cross between P1 and P2 can only be 0464 According to the present invention, it is possible to achieved by induction of staminate flowers on the pistillate apply dosage data to creating custom blended granular mixes plants by an exogenous application of the chemical silver for rolled delivery, pellets for bowls and house one-hitters, thiosulfate. This process allows otherwise pistillate (female) extracts for dabs, etc. with the flowers of these highly resinous plants to be coaxed to produce staminate, pollen bearing newly-developed varieties with designed cannabinoid con flowers. During this process, to investigate and exclude the tent so as to reduce adverse effects associated with THC. possibility of maternally inherited genetic factors, reciprocal crosses will be made where both P1 will be induced to pro Gold Class Breeding Regime duce pollen and fertilized P2 (Lina 1A), and P2 will induced to produce pollen and fertilize P1 (Line 1B). The resultant 0465 Basic Breeding Scheme. The initial cross for the F1=CBD-GOLD Lines 1A, 1B. Gold Class Breeding Regime that will be conducted is as 0473. It should be noted for accuracy that P1 (GO13) is a follows: P1 (GOLD Line (GO13)xP2 (CBD Line chemotype I plant, and crossing it to P2 (CBD Line (CBD1201). The hybrid cross between Parent 1 (P1) and (CBD1201), a chemotype III plant, the resulting population Parent 2 (P2) could only be achieved by induction of stami will yield all Chemotype II plants of intermediate THC and nate flowers on the pistillate plants by an exogenous applica CBD levels, in agreement with de Meijer, E. P. Bagatta, M. tion of the chemical silver thiosulfate. This process allows otherwise pistillate (female) plants to be coaxed to produce Carboni, A, Crucitti, P. Moliterni, V M, Ranalli, P. Man staminate, pollen bearing flowers. During this process, to dolino, G (2003) The inheritance of chemical phenotype in investigate and exclude the possibility of maternally inherited Cannabis sativa L. Genetics 163(1):335-346. 0474 Individuals from the F1 lines will be analyzed via genetic factors, reciprocal crosses will be made where both GC/MS to determine their respective chemotypes. In accor P1 will be induced to produce pollen and fertilize P2 (Line dance with de Meijer (Id) it is expected that the F1 popula 1A), and P2 will be induced to produce pollen and fertilize tions will comprise individuals that show a Chemotype II cannabinoid distribution, with both intermediate levels of P1 (Line 1B). tetrahydrocannabinol (THC) and cannabidiol (CBD). Plants 0466. These crosses will result in the production of two F1 with suitable terpene contents and profiles from this popula populations —CBD-GOLD Lines 1A, 1B. Individuals from tion will be backcrossed to the P1 parent to reinforce the the F1 lines of each F1 population will be analyzed via genetic background of P1; this population will be labelled GC/MS to determine their respective chemotypes. It is BC11: (Backcross 1 to P1). expected that the F1 populations will comprise individuals 0475 Plants from BC11 will be cannabinoid chemotype I that show a Chemotype II cannabinoid distribution, with and chemotype II; chemotype I plants will be discarded, and intermediate levels of both tetrahydrocannabinol (THC) and chemotype II plants approaching the desired terpene profile cannabidiol (CBD). will be self crossed to yield an F2 population. The BC 11 F2 0467 Plants with suitable terpene contents and profiles population will be grown, and again analyzed via GC/MS to will be self-fertilized to create a series F2 segregating popu reveal the chemotype of the plants in the population. This lations or families; all non-desirable lines will be rejected population will yield 25% chemotype I plants, 50% chemo from the breeding regimen. In this way, a series of F2's will be type II plants, and 25% chemotype III plants; only chemotype created=1 AF2a, 1 AF2b, 1 AF2c, 1 BF2a, 1 BF2b, 1 BF2c, etc. III plants will be kept for further terpene analysis, all others 0468 F2 families will be propagated and screened via will be discarded. Suitable chemotype III plants approaching GC/MS to determine individual chemotypes; it is expected the desired terpene profile will be mated via a backcross that in the F2 segregating populations we will see chemotype scheme to the P1 plant (GOLD Line (GO13)). This popula I, chemotype II, and chemotype III plants. Chemotype I and II tion will be labeled BC21; Backcross 2 to P1). plants will be discarded and only chemotype III plants will be 0476 BC21 will produce chemotype II plants. Individuals retained and again screened by GC/MS to evaluate their suit will be screened for desired terpene profile and content, and ability in terms of terpene content and profile. desirable plants will again be self-fertilized, to yield a popu 0469. These selected chemotype III plants can be self fer lation producing plants in the expected ratio of 25% chemo tilized or mated inter se and will retain the chemotype III type I, 50% chemotype II, and 25% chemotype III. Chemo cannabinoid profile within the population. It may also be type III plants will be retained for further terpene profiling desirable to mate selected F2 lines via a backcross scheme to and analysis, all other plants will be discarded. After terpene the P1 GO13 to reinforce the GOLD genetic background, analysis, selected plants from the chemotype III individuals although doing so will re-introduce B(t) alleles (i.e., the alle will again be backcrossed to the P1 plant (GOLD Line les that encode for THC production) into the breeding popu (GO13)). This population will be labeled BC31; Backcross 3 lation, resulting in a population of chemotype II plants which to P1). US 2014/029851 1 A1 Oct. 2, 2014 37
0477. From this mating, the resulting population will be 0484. In one non-limiting example, the seeds and plants comprised of only chemotype II plants, and it is expected that are implanted with a very small active RFID tag or chip which plants of the desired terpene profile will be isolated after will emit a unique address for each seed and/or plant to a analysis for terpene profile and content. reader. RFID is a wireless data collection technology that uses electronic tags for storing Substantial amounts of data that can Silver Class Breeding Regime be used for tracking individual items. There are two basic types of RFID tags: passive and active. “Passive' tags have no 0478 Using the procedures described above for the Gold power source but use the electromagnetic waves from a reader Class Breeding Regime, the initial cross for the Silver Class (e.g., the receiver) up to approximately 15 feet away to trans Breeding Regime will be as follows: P1 SILVER Line mit back their contents. Active' tags use a battery to transmit (SIL4)xP2 CBD Line (CBD1201). The basic and backcross up to about 1,500 feet. The MID tags are read when they are selection and breeding procedures for this class are as within the proximity of two-way radio transmitter-receivers, described above for the Gold Class Breeding Regime. or readers, which send a signal to the tag and read its response. The handheld devices can easily be used to track the REID Green Class Breeding Regime tags integrated into the cannabis seeds, plants, and/or prod 0479. Using the procedures described above for the Gold uct. Class Breeding Regime, the initial cross for the Green Class 0485 Alternatively, the medical cannabis plants can be Breeding Regime will be as follows: P1 GREEN Line tagged by recombinantly engineering them to express a phe (GRE1)xP2CBD Line (CBD1201). The basic and backcross notypic trait unique to the strain. For example, a strain can be ing selection and breeding procedures for this class are as stably transformed to express bio-markers, generally pro described above for the Gold Class Breeding Regime. teins, that directly, or on contact with Suitable Substrates, yield a characteristic color, optical density, light emission, or Yellow Class Breeding Regime fluorescence. Fluorescent bio-markers can include green fluorescent protein, red fluorescent protein, yellow fluores 0480. Using the procedures described above for the Gold cent protein, blue fluorescent protein, or variants thereofthat, Class Breeding Regime, the initial cross for the Yellow Class when expressed, will emit a color under a particular wave Breeding Regime will be as follows: P1 YELLOW Line length. Detection devices for fluorescent bio-markers can (YEL3)xP2CBD Line (CBD1201). The basic and backcross have one or more excitation light sources for emitting light of ing selection and breeding procedures for this class are as a wavelength or a range of wavelengths suitable for inducing described above for the Gold Class Breeding Regime. the fluorescence. In a non-limiting example, an expression cassette comprising red fluorescent protein is stably trans Purple Class Breeding Regime formed into the plant cells using standard laboratory tech 0481. Using the procedures described above for the Gold niques. This protein will be expressed by the seed and/or Class Breeding Regime, the initial cross for the Purple Class plant, and when excited by a particular wavelength produced Breeding Regime will be as follows: P1 PURPLE Line by a simple device. Such as a hand-held light, can be easily (PUR3)xP2CBD Line (CBD1201). The basic and backcross identified by the red color. ing selection and breeding procedures for this class are as described above for the Gold Class Breeding Regime. Example 4 Example 3 CBD Enhancement Dosage Technologies 0486 While it is beneficial to increase the ratio of CBD: Tracking of Cannabis Plants During Production, THC in medical cannabis, this typically is accompanied by a Processing and Use loss of the terpenoid constituents responsible for the aroma 0482 Medical cannabis must be easily distinguished from and flavor found in high THC cannabis. We can apply the recreational cannabis and hemp, allowing it to be tracked optimum CBD:THC ratios discovered from studying the from seed to plant to processing to sale (“seed to sale” track effects of the inhaled hybrids to blends, pellets, plugs, edibles, ing). This can be accomplished by tagging the seeds, har and all other medical cannabis products. The cannabis com vested material, and marketed product in a variety of different positions and dosage technologies disclosed herein can be ways. According to the present invention it is possible to used to improve the dosage consistency and Standards of raw provide instantaneously the use of forensic-style forensic flowers, blends, hashes or any other cannabis and cannabis audit capabilities to indoor horticulture. For example, the containing products provided by the present invention. compositions and methods of the present invention can be 0487 Blends comprising the cannabis strains of the used to track medical cannabis plants, medical cannabis plant present invention can be made by combining the plant mate parts, ground medical cannabis plant material, compressed rial from a high CBD:THC strain in amounts calculated to medical cannabis plant material, etc. Thus, according to the deliver the proper dose of each compound to the patient with present invention, one can track the chemotype for an indi the plant material from a strain comprising high amounts of vidual plant or group of plants from seed to flower and the desired terpenoids. The final blend contains CBD and beyond. THC in the optimum dose described herein and also an 0483 First, the seeds and plants may be implanted with a amount of terpenoids adequate to provide a pleasant flavor tracking device, such as via radio-frequency identification and taste. (RFID) using an RFID tag or chip, a telometric thread, a 0488 Alternatively, purified terpenoid extracts can be microchip, or a magnetic tag, which will allow real-time added to the plant matter from a high CBD:THC strain in a identification of the seed, plant, harvest, or final product. concentration so that the final product contains CBD and US 2014/029851 1 A1 Oct. 2, 2014
THC in the optimum dose and an amount of terpenoids (See, Code of Federal Regulations, Title 21, Volume 5, Apr. 1, adequate to provide a pleasant flavor and taste. 2009). However, the formulations studied were oral formula 0489 Finally, a purified cannabinoid extract, such as CBD tions comprising terpin hydrate, not vaporized, inhaled terpin extract, can be added directly to the plant matter from a hydrate which may prove more effective. cannabis strain comprising the desired terpenoids so that the final product contains CBD and THC in the optimum ratio/ Example 7 dose and an amount of terpenoids adequate to provide a pleasant flavor and taste. Pelletization of Blended Ground Flowers for Bowls 0490 These compositions can be used to make any type of and Pipes desired medical cannabis product including, but not limited 0497 CBD Enhancement Dosage Technologies were used to, pellets, plugs, and edibles. to create medical pre-pressed bowls of blended and pelletized Example 5 cannabis. Novel design and pellet density were used to opti mize dosage for vapor and combusted cannabinoid delivery. Blended Bubble Pack Doses The purpose of this invention is to maximize the exposed Surface area of the pelletized material to maximize contact 0491. It is important that the medical cannabis of the with heated air to achieve optimal vaporization. The preferred present invention be stable and possesses a long shelf-life embodiment is likely a very thin coin shape. when prepared for distribution to patients. This is achieved through proper drying and curing of the processed medical Example 8 cannabis product. However, the shelf-life can be increased by proper airtight packaging Such as in a bubble pack or ablister Plugs for Vaporization Technique (Specifically pack. Dabbing) 0492. The longevity (i.e., shelf-life) of the packaged can nabis can be further extended by Modified Atmosphere Pack 0498 CBD Enhancement Dosage Technologies were used aging (MAP), a technique used for prolonging the shelf-life to create medical pre-pressed gelatin-like balls for hot nail of fresh or minimally processed foods. In this preservation vaporization. The present invention provides novel designs technique, the air Surrounding the product in the package is and pellet density that is optimized for dosage and vaporiza removed by vacuum or modified to contain different levels of tion in pens and dab rigs. The purpose of this invention is to nitrogen, oxygen, and carbon dioxide. provide a precision pre-packed dose of cannabis oil for vapor 0493. The cannabis products of the present invention, ization in an e-cigarette type device or using a hybridglass or including the blended cannabis compositions described metal pipe that has been modified to operate at below com herein, can be packaged in a bubble pack in either multi- or bustion temperature levels. single-dose units to increase product longevity. Each single dose unit packaged in the bubble pack will comprise the Example 9 optimum CBD:THC dose identified by the instant invention. In one embodiment, the compositions of the invention are Horticultural Practice (Consistency) packaged as single-dose units to ensure the patient receives a 0499 All cannabis germplasm and cuttings of cannabis correct, standardized dose and to protect the product integrity. germplasm are established in identical environmental condi tions (-80° C., 80% Humidity, CO2 variable, 3000 k light Example 6 ing). Once roots are established, plants are transplanted into 1 gallon pots using a proprietary soil mix #1 heavily laded with Use of the Invention as Expectorant beneficial microbes, nematodes and predator mites. Our soil 0494. When vaporized and inhaled, the medical cannabis system is crucial to establish consistent growth patterns and varieties of the present invention are an effective expectorant. secondary metabolite production. Use of the high CBD containing cannabis varieties described (0500 Plants are grown under 18 hours of light with 50% herein can be used, for example, in the treatment of conges Metal Halide & 50% High Pressure Sodium Light bulbs tion and upper respiratory diseases. generating the spectrum. The environmental conditions, dis 0495 One mechanism through which medical cannabis tance from light, pots and Soil are all proprietary. may act as an expectorant is through the activity of terpin 0501. Once roots are bound, or plants are approximately hydrate, a precursor to terpineol which has been identified in 12"-18", they are transplanted into 3 gallon pots with propri several cannabis strains (See, Ross and ElSohly, (1996). J. etary soil mix #2. Again, microbial content of soil and ben Nat. Prod. 59:49-51 and Fischedicket al., (2010) Phytochem eficials are a crucial contributor to the consistent production istry 71:2058-2073). The presence of terpineol, instead of of medicine. terpin hydrate, in the samples after the cannabis is dried and 0502 Plants are induced into flowering by undergoing a heated may be due to a dehydration reaction of terpin hydrate period of 72 hours of darkness which is followed by the light to terpineol under thermal conditions. This chemical process cycle of 12 hours of light and 12 hours of dark (20% Metal may not occur if the cannabis is exposed to the lower heat of Halide and 80% High Pressure Sodium). Plants are trimmed, a vaporizer. pruned and topped similar to fruit tree industry (I.e., a healthy 0496 Inhalation of the vapors produced by high CBD number of budding sites distributed evenly throughout the containing medical cannabis exposed to a lower heat can act canopy). The specific techniques employed are cultivar spe as an effective expectorant and can be useful in the treatment cific. of congestion. Terpin hydrate was commonly used in the 0503 Environmental conditions, pots, distance from light, treatment of acute and chronic bronchitis, but it was removed trellising techniques, carbon dioxide concentration and nutri from the market by the FDA, which cited a lack of efficacy ent regimen are all proprietary. US 2014/029851 1 A1 Oct. 2, 2014 39
0504 Flowering period can last between fifty and ninety 0510. In some embodiments, the sensors can include sen days. While plants can exceed 5' in height, canopies are sors for air particulate/contamination measurements. In some shaped in row crop tradition and kept at 18"-24". embodiments, the sensor(s) includes a Thermo Scientific 0505 Plants are culled if they are showing expressing TEOM 1405 continuous particulate monitor. In some stress genes and/or if they are showing any signs of variations. embodiments, the air sensor(s) can include environmental Ripeness is specifically determined by genetics. controllers having sensors associated therewith, such as the Sentinel CHHC-4 that measures, in real time, temperature, Example 10 relative humidity, and carbon dioxide content. In Such embodiments, the controller can also be employed for envi Feedback-Based Cultivation System ronmental control. For example, the CHHC-4s ability to hold a set point within a certain range of accuracy can be exploited. 0506 Some embodiments of the present invention are 0511. In some embodiments, water and/or fertigation directed to systems, apparatuses, and methods for feedback parameters can be measured by a variety of sensors, including based cultivation of the herbal medicines described herein. pH, EC, flow rate, TDS, NPK, ppm of certain compounds, 0507 FIG. 1 illustrates a system 100 for feedback-based and/or others if desired. Some of these parameters can be cultivation of the herbal medicines described herein, accord determined via direct measurements, while other, such as ing to some embodiments. The system 100 includes at least a ppm of Some compounds, can be determined via dilution computing apparatus 102, an environment management sys calculations. In some embodiments, water and/or fertigation tem 104, and a patient management system 106. The various parameters can be controlled using systems such as, but not components of the system 100 can be in communication as limited to, the Hanna Instruments computerized fertigation indicated by lines in FIG. 1 via a network (wherein a dotted system (Model HI 10000) that allows for mixing of four line indicates an optional connection), which may be any type nutrient Zones and one acid/buffer Zone for pH control, and of network (e.g., a local area network or LAN, a wide area uses reliable and accurate Dosatron D8R venturi style injec network or WAN, a virtual network, a telecommunications tors. The HI 10000 can also be hooked to a reservoir style network, the internet and/or the like) implemented as a wired system or in-line flow mixing, where the preferred method is network and/or a wireless network. Any or all communica likely reservoir for compost teas and inline for fertigation. tions may be secured (e.g., encrypted) or unsecured, as is 0512. In some embodiments, the environment manage known in the art. ment system 104 can be configured to track active ingredients 0508. The environment management system 104 can be from their concentrations on the plant in the field, through configured for production of the herbal medicines disclosed production and processing. In some embodiments, the envi herein. In some embodiments, the environment management ronment management system 104 can be configured to mea system 104 can be configured for managing a controlled Sure the production of key secondary metabolites and/or environment for production of the herbal medicines disclosed monitor their flux in concentration over time to better under herein. The controlled environment can include one or more stand and control the mechanisms underlying their biosyn Software and/or hardware components monitored and/or con thesis. In this manner, aspects of the environment manage trolled by the environment management system 104 includ ment system 104 overcome challenges associated with the ing, but not limited to, one or more sensors, one or more production of herbal medicines that have multiple active controllers, one or more fertigation systems, and/or the like. ingredients, where consistent production of these active For example, in some embodiments, the environment man ingredients typically varies from crop to crop. Additional agement system 104 can include controlled environment benefits are realized when a highly monitored controlled cul grow rooms, sensors, fertigation devices, and further com tivation environment can be utilized in conjunction with puter networks and interfaces for monitoring/control of these timely chemical fertilizers that trigger the plants to produce aspects. In this manner, the disclosed embodiments are con these metabolites at the desired concentration. As a result, figurable to implement a Smart grow room, where sensor harvesting at the optimal time can guarantee consistent medi technology and artificial intelligence-based software com cine. In some embodiments, the environment management bine to assist cultivators to monitor the dozens of parameters system 104 can be further configured to optimize for indi that must be optimized to grow the highest quality and vidual metabolites of interest with troubleshooting mecha healthiest plants producing consistent levels of secondary nisms to identify issues before they impact a plants primary metabolites (as will be described in more detail later). In some or secondary metabolite production. embodiments, the sensors can include soil sensors for taking In some embodiments, the environment management system soil measurements such as, but not limited to, soil moisture, 104 can be structured in a multi tier manner and particularly electrical conductivity (EC), available soil moisture, potential in a three tier manner, with the primary order being a central gravity, temperature, and/or the like. control center/database, second order being an on-site pc 0509. In some embodiments, where grow rooms are interface station, and third order being an individual station employed, multiple sensors per room can be employed. For Such as a tablet interface. The data processing and analysis example, the total density or number of sensors in each cell can be carried out by the more powerful control center com (or room with five 4'x16' rows, ~150 plants, 15 plants per puters, which can be equipped with the latest microcomputer 4"x8" table) can vary from 2-4 per room. The number of needed forbidirectional data transmission, allowing them to sensors in a room can be dictated by the density of plants in communicate with the on-site PC stations and/or to the indi each table. Two sensors are needed for each density, whether vidual stations. The bidirectional data transmission between it is 15 or 21 plants per table, one on a boundary plant and one different facets of the network, such as the individual stations on a middle plant. Additional pairs of sensors can be added for and on-site PCs, can be accomplished in the manner outlined a specific cultivar if it is known to have substantially different in FIG. 2, which illustrates an exemplary and non-limiting water usage than Surrounding plants in the cell. embodiment of the environment management system 104. US 2014/029851 1 A1 Oct. 2, 2014 40
0513. As illustrated in FIG. 2, environmental sensors (“ac action; assign patient ID automatically and associate that ID tuator) sense environmental parameters and take in raw data with all future activities related to the patient; swipe driver's (“data') from their respective system and location therein. license upon Subsequent visits—Swipe can bring up patients This data is then location and time stamped and sent to the record and enable dispensary staffer to immediately see on-site PC station ("on-site PC). “attached scans of physician recommendation, photo ID as 0514. The raw sensor data can then be received at the well as recommendation expiration date; recommendation on-site PC. Decision making data analysis may be done on the date can be color coded to quickly draw attention if out-of on-site PC, and/or at the central control center (“central PC), date or if within X days of being out of date so that dispensary and/or other network computers as well. The data received at staffer can inform patient on the spot that either the recom the central PC is sent to the control center, and changes to the mendation is no longer valid or that it will be invalid in X data can be made by the on-site PC in conjunction with the days/weeks/months and that S/he should take steps to renew applicable system hardware. it; information regarding allocations to specific patient can be 0515. In some embodiments, a wireless system of sensor captured (date, variety, amount, S. lot) and accessible by staff to-PC communication can be used. In some embodiments, as by doing a patient “name query; feedback regarding prior best illustrated in FIG. 3, a wireless mesh network of sensors allocations can be captured (noted effects) and ratings of can be employed that feedback to a centralized pc system. medicines; follow up, correspondence to physicians can be The wireless system can contain at least three main compo prepared automatically by pulling data from allocation data nents; intelligent sensors/actuators, wireless mesh network of base fields; the ability to query database by age, gender, routers and gateways with intelligent routing algorithms, and strain, lot it, feedback (feedback itself and/or condition), etc. control and actuation. ... and cross reference with production fields below; and the 0516. In some embodiments, functionality and/or data ability to predict/recommend medicine based upon prior rat associated with the environment management system 104 can ings/preferences cross-referenced with Strain chemistries. include, but is not limited to, one or more of the following: 0520 System access can be a concern in such multiuser number of plants put into Veg (date, variety); assign lot and environments. Accordingly, embodiments directed to system plant number, track development—ability to ascertain Inven access will be described with respect to the system 100, and tory of plants at any given time; assign date of flowering (date unless explicitly stated otherwise, are understood to be flowering initiated, variety, lot it, plant it, location); track directed to aspects of operation of the environment manage feeding schedule during flowering (date, six nutrient fields); ment system 104 (also referred to as the production side), track environmental conditions (linked to various sensors in and/or the patient management system 106 (also referred to as the room: Soil moisture, temperature, humidity, CO level. the patient side), and/or the computing apparatus 102. In and Light intensity); cultivator notes field (Date, Note field Some embodiments, system access (production side and for cultivator to make notes on specified date, i.e., lights were patient side) can include four components of hierarchy; mas mistakenly left on form 0200 until 2300); cannabinoid/ter ter administrator, regional manager, on-site manager, and penoid testing log (results, testing date, point in flowering); cultivator (production side)/counselor (patient side). Com harvest (date, B&T weight); processing (trim date, weights); munication structure can be cloned from one tier to the next, bulk packaging; transit; acquisition from MPC lot #, Vari e.g. from cultivators to master administrators. In some ety, production reimbursement, total weight, form; receipt embodiments, the communication structure can include (entity, name, date); safety screening results (pass/fail)— alerts, decision tree confirmations, and/or other clearance molds, pesticides, aflatoxins, microbial; weighing; assembly restrictions—most restrictive at the cultivator level and least (units); allocation information (amount, avg. allocation, at the master control level. This overlapping of communi reimbursement); and popularity indices (rank, Velocity, cation in each sector can bring continuity between the chain potency/reimbursement via cross-references with “Patient’ of command so that major decisions are always cleared on data). Tables 23 and 24 illustrate exemplary and non-limiting multiple levels. Integrating with the on-site PC and individual embodiments of the cultivation-related information that can PC can condition operation patients to use open communica be collected. tion that they know is backed by System-checked account 0517 Referring again to FIG. 1, the patient management ability. system 106 can be configured to acquire patient data in any 0521. The on-side PCs and the individual PCs can have a Suitable manner. In some embodiments, the patient manage private communication system therebetween, such as ment system 106 can be configured to record patient data encrypted IM and/or some form of closed/private network. In within the context of a method as illustrated in FIG. 1. Some embodiments, emails are encrypted for patients that can 0518 Tables 21 and 22 illustrate exemplary and non-lim send notifications to users’ email of choice when a new email iting embodiments of the patient-related information that can arrives in their encrypted box. be collected, including prescribing physician information. 0519 In some embodiments, functionality and/or data PC Computer Terminal Interface associated with the patient management system 106 can Master Administrator Platform: include, but is not limited to, one or more of the following: standardized and compliance messaging to visitors (clients, 0522. In some embodiments, the master administrator elected officials, healthcare providers and media) by using platform (e.g. the central PC) can be characterized in the recorded images/messages transmitted electronically via tab following exemplary and nonlimiting manner: access to all let (this can include all agreements and consents); collect real-time databases, archived data sets/analysis results, biographical, contact, health history, and prior non-cannabis patient information, cameras, etc. No access restrictions, treatments electronically (currently collected on handwritten access can be heavily encrypted and access codes can be very forms); set up patient record automatically; immediately limited in number, only to key company patients for example. upon completion of registration process, prior to first trans Access to certain aspects of the master platform can be par US 2014/029851 1 A1 Oct. 2, 2014
titioned off for limited access to other manager(s) if needed. represent changeable variables in the at-a-glance Screen). For example, lab managers can have access to analysis data, Any analysis done by algorithms could also be accessible at certain production managers have access to some production the individual PC level, but not necessarily as in depth as is data, etc. available at the on-site PC level. In some embodiments, the master access platform includes the ability to make changes/ Regional Manager Platform overrides that update to selected individual PCs (i.e. a master 0523. In some embodiments, the regional manager plat access change to nighttime temperature schedule for a certain form allows for control over a number of sites, and over cell would send notifications of the change, if desired, and selected parameters that can be delegated by the master create a permanent change). In some embodiments, the mas administrator platform. For example, the person responsible ter access parameter set points, and other system parameter for formulating fertigation Solutions in the lab can have elements that are outside of the regional manager/site manger regional access over the fertigation/soil water parameters, but security clearance will require an encrypted password to change. If needed, this would allow master administrators to not full master access to all sites. This access can be grant lower management access to certain elements on per restricted further to be allowed from on-site network comput case basis. CS. 0529. In some embodiments, the master access platform Site Manager Platform includes the ability to access patient records, Surveys, Survey group data, blood sample data, and all other aspects of the 0524. In some embodiments, the site manager platform patient side of the system. At-a-glace home screen for patient (e.g. the on-site PC) provides an access point for data com data will have the ability to show output of algorithmic data pilation/entry, Excel, Word, system specific software, and/or mining. A patient system example would be when a patients the like. System access/control will encompass control over master filtered parameters such as fertigation time series/ makes their first visit and submits their information into the allocation and/or the like. Any independent changes made by patient database, that information is cross-referenced with an the site manager either via their individual PC interface would array of other patient data points (such as ailment, age, be sent back to the necessary upper management in the form gender, Survey responses, chemovar preference, etc.). Based of an email, IM, and/or other chosen alert method. In some on the results of one or a few simple data mining algorithms, embodiments, a two method minimum, and preferably three tailored recommendations can be made and generated on the alert methods are preferred for adequate redundancy and counselor's individual PC in real time (e.g. a recommenda accountability. tion engine can be implemented). 0525. In some embodiments, no cultivator/counselor 0530. In some embodiments, the master administrator access is permitted to on-site PCs or otherwise, and cultivator platform can include the ability to set the recommendation interaction can take place through the individual PC only. parameters for the algorithms decision process, but whatever Counselor access will take place through a separate indi chemovar recommendation parameters are chosen, in some vidual PC intended to provide product information to inform embodiments, they can remain constant for all new patients. counselors and, through the counselors, consumers. In this manner, a consistent reliable database can be built over time, which will increase the accuracy of the system. This Individual PC Platform ability for the system to learn using AI (artificial intelli gence) software programming, likely with evolutionary algo 0526. The individual PC will serve different needs for rithms, will require a certain amount of time of patient different levels of management and operators, but the main response data to be entered before the programs(s) can dis purpose can be for use as a company specific interface and cern which decision pattern yields the favorable result a sta communication tool. At all levels users can populate, manage, tistically significant amount of times. The eventual result of and track their tasks, as well as enter data and notes. In some this system component at the patient/counselor interaction embodiments, all users can also send and receive messages to level can be an accountable and consistent decision tree pro other users within their realm. At higher levels, users can cess that is tied in to all levels of management, removing track data trends, view real time data, and analyze various counselor recommendation variance from one to the other data components in different graph formats and analysis and possible misinformation. Although this example pertains methods of their choosing. This analysis will tap data on the to chemovar recommendation, it is understood that it can also master database for all sites, allowing regional manager and be applied to other patient/counselor interactions such as master administrators to track multiple site data from one patient/POA (point of allocation) and others. device. 0527 The level of interaction at each level can happen via 0531 Regional Manager Platform: The individual PC applications in Some embodiments, some shared by all users regional manager platform can allow RMs to have at-a-glance and others only for those with special permissions. A majority data viewing/comparison capabilities similar in function to of these applications can be specific need-based adaptations that of the master administrator, but restricted in content to of preexisting native apps (i.e.: notepad) or proprietary apps. that which is job/project related or delegated otherwise. Data 0528 Master Access platform: Data input for all areas of input/analysis and system monitoring can be the main use of production and/or patient side. In some embodiments, the the individual PC for RMs. Selective control over certain master access platform allows viewing of each site(s) critical master delegated system parameters could be altered by data at a glance. The at-a-glance data can be changed in RMs via the individual PCs similar to the way it would be on both content and form. For example, one patient may want to the on-site PC, but via a comparatively deconstructed/re compile yield data for all sites that are displayed in a monthly fined tablet interface. time series linear graphs over a prior year, with a year-to-date 0532. Site Manager Platform: Can allow for site overview production trend graph for comparison (underlined portions and management of multiple cultivators or cultivation teams. US 2014/029851 1 A1 Oct. 2, 2014 42
0533. Cultivator/Counselor platform: Can allows for cul (0544 Chemotype Profiles tivator notes to be entered into the system, and the system can 0545. These can have the same framework as the terpene digitally tag the notes with date, time, batch number, plant program, but can include cannabinoids and other secondary number, etc. in the system to be referenced at a later point if metabolites of interest. needed. Cultivators will need to have fields in the notation 0546 Bioinformatics application that will be filled out with the appropriate infor 0547. The use of evolutionary algorithms to run computer mation to create a track record for the entry tag. models of mass breeding programs that can allow for 0534 Having described system access, referring again to increased efficiency in parent material selection as well as FIG. 1, embodiments directed to software tools will be accurately estimating required population sizes for field tri described with respect to the system 100, and unless explic als. itly stated otherwise, are understood to be directed to aspects 0548 Algorithms for Data, Systems and decision Making of operation of the environment management system 104 0549. Numerous algorithms can be used at any point either (also referred to as the production side), and/or the patient singularly, simultaneously or in conjunction to produce new management system 106 (also referred to as the patient side), data, maintain system functionality and/or optimization, and/or the computing apparatus 102. compilation and execution of fuzzy control programs, ana 0535 Decision Tree Analysis Help Tool lyzing and/or processing data, making system updates and 0536. Designed with the cultivator/counselor in mind, this intelligent decision/changes, and monitoring system com application can serve both as a communication pathway ponents/sensors to name a few. Some of the algorithms used between managers and cultivators/counselors as well as a to address dynamic data sets and problems can include, but help tool for them as well. A troubleshooting function is in the are not limited to; least squares algorithms, direct and/or form of a series of searchable common issues that arise either indirect control evolutionary algorithms, pattern recognition in daily procedure or possibly on rare occasions. If such an algorithms, data fusion and/or data clustering algorithms. issue arises that someone doesn't know the correct flow of 0550 Referring to FIG. 1 again, the computing apparatus action for a particular task, they can reference this application 102 (also referred to as the “central computer, the “central to see a decision tree/flow chart on how it should be done PC, etc. See FIGS. 2 and 3) can handle the acquisition, according to management. processing, and analysis of data from different components of 0537. This application can become a communication tool the system 100, including the environment management sys when the managers, whether transitory regional or permanent tem 104 and the patient management system 106. In some on-site, choose to upload decision trees into the system. For embodiments, the computing apparatus 102 can be config example, if a regional manager comes through and makes ured to track both crop and patient trials of chemotypes of changes to operating procedure or wants to reiterate proce potential interest. For example, the computing apparatus 102 dure, they can quickly create a simple decision tree chart can be configured to track the production of metabolites of (possibly pre-formatted entry fields) while on-site and upload interest in a crop, while also being configured to track the that system onto the network. Once uploaded, it is available metabolism of those eventual plant-produced metabolites as for others to view when needed, and managers could even they are metabolized by consumers. Thus, active ingredients make it into a checklist format in which operators must check can be tracked from their concentrations on the plant in the off steps in the process until proficient. field, through production and processing, to the eventual con centrations as metabolites in the blood of patients, post con 0538 Data Entry Portal Sumption. In this manner, aspects of operation of the comput 0539. The data entry portal can be the data entry applica ing apparatus 102 can define the complete chemical tion for the individual PC that will have different forms for relationship between plant and human. In some embodi different operator positions. For example, patient-based entry ments, this defined chemical relationship can be used to cre fields (i.e.: POA data, patient feedback data, etc.) for counse ate maps, multi-dimensional scatter plot to examine and/or lors and plant-based entry fields (i.e.: plant number, lot num analyze patterns within a host of metabolic variables through ber, package number, etc.) for cultivators. out the incredibly complex system. (0540 Data Analysis Tool 0551. In some embodiments, once data is received at the 0541. The data analysis tool can allow managers and tech computing apparatus 102 any number of actions can be taken, nicians the ability to alter their at-a-glance home screens and based on a user's needs and based on a user's associated run other analysis on their data in the field. The range of this system access parameters as discussed above (i.e. a user of the analysis can be limited in comparison to the pc interface. The computing apparatus 102, of the environment management results of Such an informatics system can be directed and system 104, and/or of the patient management system 106). displayed in many ways, to be chosen by the user. In some embodiments, the computing apparatus 102 can be 0542 Genetics Terpene Profiles configured to implement one or more algorithms to analyze 0543 System is designed to analyze, characterize and various types and forms of information including, but not codify the subtleties in terpene differences across a large limited to; genetic data, breeding data, tissue culture data, number of separate genetic groups (as per the color coded field trial data, all computer system-related data, greenhouse system), different populations within those groups, and time data, indoor grow data, environmental sensor-sourced data, series analysis tracking where applicable (i.e.: terpene ratio environmental data from other sources, all patient-related/ and/or quantity variation during final weeks of flower devel Sourced data, allocation/reimbursement data, and all other opment). Individuals will be grouped into different color types/forms of proprietary sourced data. groups based initially on some qualitative characteristics Such 0552. The resulting information can then be transmitted as nose (piney, fruity, etc.), and later quantitatively. Quan back to the user that requested it in the form of their choosing titative analysis will allow for each individual to be profiled viabidirectional data transmission. This transmission, either into the database. wireless or wired in signal, can be routed through the network US 2014/029851 1 A1 Oct. 2, 2014
(not shown), and/or can be encrypted. The user can then TABLE 22 choose to make changes or updates to the controllable/acces sible aspects of the system 100, if applicable. For any alter Exemplary Recommending Physician Table ations to system parameters or any other significant system recommending physician last name (relates to Patient Table) aspect, a feedback system can exist for alerts, timestamps, recommending physician first name (relates to Patient Table) updates to current/future computational processes, refer recommending physician Street name and number recommending physician address designation: avenue? drive street/none enced data sets, and other signals. recommending patient Suite number up to 4 digits) 0553. In this manner, patient feedback data can fuel the recommending physician Zip code (5 digit) production of medicines. For example, the patient feedback recommending physician business phone (10 digit) data can be used to optimize pharmacologically active plant recommending physician business name oil content through a host of breeding and cultivation tech Condition Table(s)... niques. In some embodiments, the computing apparatus 104 can be configured to monitor market trends and identifies products appeal, efficacy, and sell-through as the products TABLE 23 chemotype evolves over time refined by consumer feedback Exemplary Lot Table and research studies. In some embodiments, the feedstock that is used to create these products can be selected in ot identifier (relates to Collective Table and Patient Table) otlocation identifier response to real-time feedback collected by this system from otplant identifier (relates to Plant Table) consumers. The coupling of chemotype development and ot date began veg selection with consumer feedback can enable the identifica ot date began flowering tion of market trends of selected chemotypes at the earliest ot feeding date(s) possible stage in product deployment. For example, principal otfeeding date(s) nutrients (six fields) ot environmental condition(s) (dates) (soil moisture, temperature, component analysis can be used to identify synergies humidity, CO2, light between groups of pharmacologically active constituents that intensity) are gaining traction with consumers for their medicinal effec cultivator notes cannabinoidterpenoid testing (results, testing date, point in flowering) tiveness, their aesthetic appeal or combination of both. Safety screening results (pass/fail) harvest date TABLE 21 harvest date weight trim date Exemplary patient table trim date weight bulk packaging date patient collective (list) (relates to Collective Table) bulk packaging date weight patient join date (XXiXX XX) transit departure date/time patient referred-by last name lant Table(s)... patient referred-by first name patient last name patient first name patient street name and number patient address designation: avenue? drive? street none TABLE 24 patient birthdate (XXXX XXXX) patient apartment number up to 4 digits) Exemplary Collective Table patient zip code (5 digit) collective lot arrival date?time patient home phone (10 digit) collective lot identifier (relates to Lot Table) patient cellphone (10 digit) collective break lot up into units date patient scanned id (yes/no) collective units inventory patient recommendation date (XX XXXXXX) patient unit allocation date patient recommendation scanned attachment (yes/no) patient unit allocation identifier patient state id card date (XX XXXXXX) patient recommending physician last name (relates to patient unit allocation reimbursement Recommending Physician Table) Seed to Plasma Constituent Analysis patient recommending physician first name (relates to Recommending Physician Table) patient consent to share data with physician (yes/no) Example 11 patient consent to share data with physician scanned attachment (yes/no) patient condition 1 Multiplexed Cannabis Medicines patient condition 1 prior treatment medication (list?) 0554. To make multiplexed cannabis medicines that are patient condition 1 prior treatment medication feedback (ranking) patient condition 2 designed to be effective in a particular disease or disorder, patient condition 2 prior treatment medication (list?) base extracts which have not undergone much manipulation patient condition 2 prior treatment medication feedback (ranking) are mixed with stock fortifying extracts that have undergone patient condition 3 extensive manipulation. These mixtures provide stocks of patient condition 3 prior treatment medication (list?) materials that are rich in either THC, CBD, CBG, limonene, patient condition 3 prior treatment medication feedback (ranking) pinene, myrcene, linalool, beta-caryophyllene, phytol, terpi patient cannabis experience (list) nolene, terpenene, ocimene, caryophyllene oxide, or alpha patient unit allocation date (XXiXX XXXX) humulene or a combination thereof. patient unit allocation date patient unit allocation date quantity (standard unit measurement?) 0555 Any means commonly used in the art to isolate patient unit allocation date feedback (Survey) (correlate to prior treatment particular cannabis agents may be used may be used to pre medication pare the fortifier stocks. For example, stock cannabinoid for tifiers with high THC (I), CBD (II), and/or CBG (IV) con patient unit allocation date feedback Survey date (XXiXX XXXX) tents, are produced by removing the extract from phenotype I. II, or IV plants that are high in THC, CBD, and/or CBG. The terpenes are distilled from the extract by supercritical extrac US 2014/029851 1 A1 Oct. 2, 2014 44 tion to provide a crude sludge, which is then winterized to This may actually serve to increase the therapeutic margin COWe WaxeS. depending on the condition. For instance, tolerance to cogni 0556. To prepare the high terpene (EO) fortifiers, plants tive and psychomotor impairment, the psychological high, are produced that have the desired concentrations ofterpenes: tachycardia, and orthostatic hypertension, tends to develop these include, but are not limited to, limonene, pinene, rather quickly and chronic users may not experience these myrcene, linalool, beta-caryophyllene, and/or phytol. The detrimental side effects, while still benefitting from the anal high terpene extract is removed from the plants, and is then gesic or other therapeutic effects of cannabis. Conversely, the steam distilled to provide stock terpene fortifiers with high novice user who has no tolerance, can be slowly subjected to limonene (L), pinene (P), myrcene (M), Linalool (LO), caryo dose escalation (e.g. over 30 days or more) to build tolerance phyllene (C), and/or phytol (P). Since these compounds may to these effects before given therapeutic doses. Many times extract together it might be necessary to fractionally distill the the dysphoria experienced by naive users is enough to cause crude to further enrich for the desired compound. discontinuation of the treatment, and slow dose escalation 0557. The cannabis base to which the high cannabinoid or which helps induce tolerance to the detrimental side effects terpene fortified stock is added is prepared from any and all of may alleviate this. the various strains described herein, or others known in the 0561. The biodistribution and PK of the cannabis active art, by supercritical extraction. This provides the foundation agents administered either orally or through inhalation differ cannabinoid ratio which retains the subjective qualities of the Substantially. An acute condition may respond better to an strain since all the cannabinoids, terpenes, and waxes are still inhaled formulation while a chronic condition may respond present. better to the prolonged plasma concentrations resulting from 0558. The concentrations of the various active agents oral administration. The higher levels of 11-OH-THC (and present in the multiplexed cannabis medicine will vary presumably CBD) formed from first-pass metabolism after depending on what has been determined to be the optimal oral formulation administration, which is more potent and has dosing for any particular disease or disorder being treated. better blood brain barrier penetration than the parent com Depending on the condition being treated and the Subjective pound, has implications for neurological conditions. The dos qualities desired, the base is then fortified with high cannab ing studies described herein evaluate the effects of various inoid and/or terpene Stock to give the final preparation doses of the multiplexed cannabis formulations when admin according to the FIG. 4. istered either orally or through inhalation. 0559 Examples of some mixtures that have therapeutic benefit are OG-(I)L (OG with THC and limonene fortifiers), Formulations OG-(DLP (OG with THC, limonene, and pinene fortifiers), 0562. The amounts and types of bases, cannabinoid and OMRA-(II)C (OMRA with CBD and caryophyllene fortifi terpene fortifiers are designed to have a synergistic effect on ers), and JJXA-(IV)Lo (JJXA with CBG and linalool fortifi the conditions being treated. The multiplexed signaling ers). resulting from the synergy of the components may be more effective than any single component alone and are tailored to Example 12 achieve the desired effects. For instance, analgesia has been shown to be mediated by the CB, CB, TRPV-1, and C-AR Dosing Regimens of Multiplexed Cannabis receptors, which Suggests a component mixture of THC Medicines (which acts on CB and CB), TRPV-1 (which acts on CBD), CBG (which acts on C-AR) and B-myrcene (which acts on Patients C-AR) will be therapeutic. Similarly if the cause of the pain 0560 Regardless of the condition being treated, two sepa is inflammation, which is mediated by TNF-C. and PGE-1, rate groups of patients are evaluated: one composed of novice then the synergistic effects of a multiplexed medicine com cannabis users and one composed of experienced cannabis prising CBD-rich extract, which counteracts TNF-C. and users. It is important to know the past cannabis use history of C-pinene, which counteracts PGE-1, proves a more effective patients since tolerance can occur in experienced users, who therapeutic than extracts not containing both of these com will therefore experience the therapeutic effects of the mul pounds. The following Table 25 shows a few examples of the tiplexed cannabis formulation differently than those with no various clinical indications that are treated with cannabis tolerance. However the rate and duration of tolerance varies formulations, the cannabinoids and terpenoids that are effec with the different effects; a particular individual may have tive therapeutics for each clinical indication, and the path developed tolerance to one cannabis agent but not to another. ways each cannabinoid influences. US 2014/029851 1 A1 Oct. 2, 2014 45
/9HT8IVIL
OHL
SW US 2014/029851 1 A1 Oct. 2, 2014 46
AOHL ZATHI(+) NI?TO
OHL US 2014/029851 1 A1 Oct. 2, 2014 47
0563 The fortifiers of the present invention are chosen to Study 1: Precision of Dosing Regimens and Bioavailability reinforce the treatment for the given clinical condition and to 0568 Traditionally, administration of cannabis has posses an improved therapeutic margin, through synergy of resulted in unpredictable bioavailabilities, resulting in fre the various pathways involved in the disease or disorder. The quent occurrences of overdosing and/or underdosing which above Table 25 is a very brief, and by no means complete, make it difficult to attain therapeutic blood levels while miti Summary of pharmacological effects of the various cannab gating adverse events in a predictable manner. Therefore, the inoids and terpenoids along with the relevant therapeutic ability to provide predictable and consistent blood plasma applications. In cases where a mechanism has been proposed levels has great utility. this has been included in the table. 0569. In this study, patients are divided into two groups: one receiving inhaled cannabis formulations, and one receiv Patient Sub-Groups and Controls ing oral cannabis formulations. Those receiving the oral dose 0564 Large patient groups (75-100 patients) are studied to of cannabis abide by strict pre-dosing dieting. The dose evaluate the subjective effects of the cannabis formulations. amount is scaled to body weight (0.05 and 0.1 mg/kg) and, For all studies, patient groups are chosen from several loca since cannabis active components are highly lipophillic, the tions and are chosen from various dispensaries and/or solic dose amount further scaled based on BMI and/or body fat ited, if drug-naive patients are difficult to find. These patients measurements. For example, the dose based on body weight are Subdivided into experienced and novice cannabis users, can be multiplied based on the patient's BMI (e.g. multiply and then if the clinical indication warrants it, further subdi ing the dose by 0.9 for BMI<18, 1.0 for BMI=18-25, 1.1 for vided into those receiving either the oral and inhaled formu BMI 25-30, and 1.3 for BMD-30). Each study is performed in lations. Due to the extremely variable bioavailability, dosage triplicate to determine intra-patient variability and each regimens are tailored to the indication and the patient. All patient first undergoes i.v. dosing with the prescribed amount studies are done with the appropriate medical and/or psycho of drug. The oral formulation is given in a single dose, and to logical Supervision and evaluation. There are several placebo minimize the effect of Smoking characteristics, the inhaled groups, with the patients receiving either complete placebos, formulation is given in tabs of sufficient size to be vaporized a placebo containing no cannabinoids and only terpenes, and and administered in a single dose. Alternatively, the tabs to be placebos containing no terpenes and only cannabinoids. This vaporized are subdivided into “unit sizes” that are adminis will serve to establish not only efficacy of the cannabinoids tered in rapid succession. Blood samples are taken at various and/or terpenes, but also the Synergy. The complete placebo is intervals and assayed for the cannabis active agent as well as generated from fats and waxes resulting from cannabinoid the appropriate metabolites. From the data biodistribution extraction and is spiked with terpenes fortifiers for exact and and appropriate PK parameters are determined. reproducible levels of terpenes to make the placebo without cannabinoids, or it is spiked with cannabinoid fortifiers to Study 2: Mitigation of Adverse Effects make exact and reproducible levels of cannabinoids without 0570. This study establishes the development of tolerance the terpenes. to the possible adverse effects of cannabis, such as cognitive and psychomotor impairment, the psychological high, anxi Proposed Clinical Indications ety, and tachycardia. This is important as many times the therapeutic dose approaches the intoxicating dose and may 0565. The studies first evaluate the predictable and repro cause discontinuation of treatment. Only inhaled formula ducible plasma levels of cannabis active agents both in a tions are employed in this study since the onset of the drug patient, and between different patients, who received the mul effect is rapid and the duration is shorter, which provides tiplexed medicines either orally or through inhalation. Once easier monitoring. Inhalation of the drug formulation is pref this is evaluated, the mitigation of adverse effects is studied erably done with a volcano, timed inhalations, timed 10-sec through dose escalation and/or examining the ratios of active ond breath-holds, and/or timed intermediate duration. Sub ingredients in the multiplexed cannabis formulation. Once jective questionnaires and heart-rate monitoring are used for this is established, the various clinical indications are exam evaluation. ined. 0571. The subjects are divided into a number of groups, 0566 Based on proposed pharmacological mechanisms of and are administered either complete placebo, placebo with action, there are a number of clinical indications that are only terpenes, THC base, THC:CBD base, or THC base with evaluated for treatment with cannabis-based medicines. varying levels and combinations of CBD, CBG, limonene, These include, but are not limited to, Parkinson's, Alzhe and/or linalool fortifiers, all of which are suggested to allevi imer's, MS, Stroke, glaucoma, anorexia, cachexia (from ate anxiety. The Subjects are administered with 3 mg, 6 mg, or AIDS, cancer, Multiple Sclerosis, congestive heart failure), 12 mg of the drug formulation (or dosage levels determined diarrhea, allergies, arthritis, irritable bowel syndrome, from Study 1). The subjects are further subdivided into those Crohn's disease, sleep disorders, epilepsy, chemotherapy who are administered the maximum dose at the first treatment induced emesis, panic disorder, social anxiety disorder, gen and those who undergo a slow dose escalation. This estab eralized anxiety disorder, post-traumatic stress disorder, lishes the proper dosing regimens and ratios of anxiolytic depression, spinal cord injury, cerebral palsy, post-operative ingredients in the multiplexed formulations if adverse events pain, migraine, neuropathic pain, Sciatica, asthma, and/or are noted in future studies. sleep-related breathing disorders. Study 3: Pain Terminology 0572 Patients are grouped into those suffering from Mul 0567. In the studies below, the medicines are referred to by tiple Sclerosis, post-operative pain, migraine, arthritis, and the principal components of the base and fortifiers. neuropathic pain (Such as Sciatica) and then Subdivided into US 2014/029851 1 A1 Oct. 2, 2014 48 those receiving either oral (2 mg, 5 mg, 10 mg, 15 mg, 20 mg Study 8: Appetite Stimulant THC) or inhaled (2 mg, 5 mg, 10 mg, 15 mg, 20 mg THC) administration routes. Dosage levels can also be determined 0577 Patients are grouped into those suffering from anor based on Study 1. Patients are administered with the placebos, exia, AIDS Wasting Syndrome, and cachexia resulting from THC base, THC/CBD base, or various amounts of CBD, MS or CHF and subdivided into those receiving either oral (2 CBG, myrcene, and/or linalool fortifiers, all of which have mg, 5 mg, 10 mg, 15 mg, 20 mg THC) or inhaled (2 mg, 5 mg. been implicated in analgesia. Patients are evaluated via ques 10 mg, 15 mg, 20 mg THC) administration routes. Dosage levels can also be determined based on Study 1. Patients are tionnaire and/or medical examination. administered either the placebos, THC base, THC/CBD base, Study 4: Anxiety limonene or pinene for associated anxiety, and CBG, or limonene for associated depression. Patients are evaluated via 0573 Patients are grouped into those suffering from gen questionnaire and/or medical examination. eralized anxiety disorder (GAD), seasonal affective disorder (SAD), panic disorder, and post-traumatic stress disorder Study 9: Neuroprotection (PTSD). Patients are subdivided into those receiving either oral (2 mg, 5 mg, 10 mg, 15 mg, 20 mg THC) or inhaled (2 mg, 0578 Patients are grouped into those suffering from mild 5 mg, 10 mg, 15 mg, 20 mg THC) administration routes. Parkinsons, Alzheimer's, Multiple Sclerosis, and possible Dosage levels can also be determined based on Study 1. recent stroke and subdivided into those receiving either oral Patients with SAD receive a lower dosing regimen. Patients (2 mg, 5 mg, 10 mg, 15 mg, 20 mg THC) or inhaled (2 mg, 5 are administered either the placebos, THC base, THC/CBD mg, 10 mg, 15 mg, 20 mg THC) administration routes. Dos base, or various amounts of CBD, CBG, limonene, and/or age levels can also be determined based on Study 1. Patients linalool fortifiers, all of which have been suggested to relieve are administered either the placebos, THC base, THC/CBD anxiety. Patients are evaluated via questionnaire and/or psy base, limonene or pinene for associated anxiety, and CBG or chological examination. limonene for associated depression. Patients are evaluated via questionnaire and/or medical examination. Study 5: Depression Study 10: Multiple Sclerosis 0574 Patients are subdivided into those receiving either oral (2.5 mg and 5 mg THC) or inhaled (2.5 mg and 5 mg (0579 Patients are subdivided into those receiving either THC) administration routes. Dosage levels can also be deter oral (2 mg, 5 mg, 10 mg, 15 mg, 20 mg THC) or inhaled (2 mg, mined based on Study 1. In this study, higher doses are not 5 mg, 10 mg, 15 mg, 20 mg THC) administration routes. examined since only low doses of cannabis have been impli Dosage levels can also be determined based on Study 1. cated in relieving depression. Patients are administered either Patients are administered either the placebos, THC base, the placebos, THC base, THC/CBD base, or various amounts THC/CBD base, or various ratios of THC fortifiers (associ of CBD, CBG, and/or limonene, all of which have been ated with neuro-protective, immunosuppressive, anti-inflam Suggested to relieve depression. Patients are evaluated via matory, anti-spasmodic, analgesic, and muscle relaxant questionnaire and/or psychological examination. effects), CBD fortifiers (associated with neuro-protective, immunosuppressive, anti-inflammatory, anti-spasmodic, and Study 6: Allergies, Rheumatoid Arthritis, Irritable Bowel analgesic effects), CBG fortifiers (associated with anti-spas Syndrome, Pain, MS, Crohn's Disease, Arthritis modic, analgesic, and muscle-relaxant effects), pinene (asso ciated with anti-inflammatory effects), myrcene (associated 0575 Patients are grouped into those suffering from aller with anti-inflammatory and analgesic effects), linalool (asso gies, rheumatoid arthritis, irritable bowel syndrome, pain, ciated with analgesic effects), and beta-caryophyllene (asso MS, Crohn's disease, and arthritis and subdivided into those ciated with anti-inflammatory effects). Patients are evaluated receiving either oral (2 mg, 5 mg, 10 mg, 15 mg, 20 mg THC) via questionnaire and/or medical examination. or inhaled (2 mg, 5 mg, 10 mg, 15 mg, 20 mg THC) admin istration routes. Dosage levels can also be determined based Study 11: Epilepsy/Migraine on Study 1. Patients are administered either the placebos, THC base, THC/CBD base, or various amounts of CBD, 0580 Patients are grouped into those suffering from sei pinene, myrcene, and/or beta-caryophyllene fortifiers, all of Zure disorders of different classifications and migraine head which have been suggested to be inhibit pro-inflammatory aches of different classifications, and subdivided into those and immune response pathways. Patients are evaluated via receiving either oral (2 mg, 5 mg, 10 mg, 15 mg, 20 mg THC) questionnaire and/or medical examination. or inhaled (2 mg, 5 mg, 10 mg, 15 mg, 20 mg THC) admin istration routes. Dosage levels can also be determined based Study 7: Asthma, Sleep Disorders, and Sleep Apnea on Study 1. Patients are administered either the placebos, 0576 Patients are grouped into those suffering from mild THC base, THC/CBD base and CBD, CBG, or linalool for asthma, central sleep apnea, and obstructive sleep apnea and tifiers, all of which are implicated in anti-epileptic pathways. Subdivided into those receiving either oral (2 mg, 5 mg, 10 Patients are evaluated via questionnaire and/or medical mg, 15 mg, 20 mg THC) or inhaled (2 mg, 5 mg, 10 mg, 15 examination. mg, 20 mg THC) administration routes. Dosage levels can Example 13 also be determined based on Study 1. Patients are adminis tered either the placebos, THC base, THC/CBD base, or Zero-Point Delivery Device and Principles various amounts of pinene, which has been implicated in bronchodillation and of myrcene and linalool, which have 0581 Vaporization is the process of heating a substance to been Suggested to be sedatives. Patients are evaluated via its boiling point to release vapor containing the active con questionnaire and/or medical examination. stituents in a gaseous state. This vapor can be inhaled to US 2014/029851 1 A1 Oct. 2, 2014 49 deliver the active agents in the drug, but without the harmful duct the high Voltage current produced by the vaporization irritants and carcinogens found in Smoke that results from device to the sludge to vaporize it. combustion of the plant material, and without the alcohol and 0588. The amount of sludge on each dosing strip is prede preserved water that serves as a base for nebulizer solutions. termined based on the patient and the disease and/or disorder There is a need for a convenient handheld and/or tabletop being treated, to provide accurate and consistent dosing. The vaporization device that vaporizes designer sludges (i.e. Solvent-free sludge is extracted via multigas extraction and material to be vaporized) that are created with predetermined comprises the refrigerant 134A, butane, iso-butane and pro and calculated resistances to work best with the vaporization pane in a ratio that delivers a very complete and balanced device. The internal resistance of the sludge, in concert with extraction of essential oils. the high Voltage current and the aluminum dosage Strip tech 0589. The predetermined quantity of sludge is applied nology described in the next Example below, generates the onto each of four or more (R-R) connections (activation necessary heat of vaporization to volatilize all of the active sites) on the dosage strip. The dosage strips are inserted into components in the sludge. a vaporization device and are activated by the device's micro 0582 The vaporization device requires, at its most basic, a chip at any number of the sludge activation sites on each source of heat that is emitted when an electric current is dosage strip. The amount of dose administered to the patient passed through a wire or a fluid, and a dosage Strip containing is selected and altered using the dosing Switch on the vapor the cannabis sludge to be vaporized that has been optimized izer. The settings on the selector switch communicate with the for consumption in the vaporization device. The design of the microchip to control how many activation sites on the strip are delivery device comprises components that are similar to that activated. of a basic taser or stun gun, which have been used in the 0590 The dosing strips and the vaporization device laboratory to vaporize cannabis oils or sludges. An example described herein, allow the cannabis active compounds to be of a sample taser design is shown. delivered to the patient in a method that is capable of repro 0583. At its simplest, the ergo-dynamic vaporization ducible and accurate dosing for essential oil and cannabinoid device described herein comprises a space for depositing the medications. dosage strip, a dose selector Switch, a micro-computer which activates any one or more four activation sites present on the Deposit Information dosing strip, an activation Switch, a battery, a speaker, a LED light, and an area through which the patient inhales the vapor. 0591. A deposit of the cannabis varieties of the present An example of one manifestation of the vaporizer device is invention, including the Classes of Cannabis Varieties, is shown in FIG. 5. maintained by the Biotech Institute, LLC 5655 Lindero Can 0584) The patient chooses the proper dosage on the dose yon Road, Suite 226, Westlake Village, Calif. 91362. selector switch, and pushes the activation switch, thereby 0592. In addition, a sample of one or more varieties of this delivering a high Voltage current through the aluminum elec invention has been or will be deposited with an International trodes on the dosage Strip to heat and vaporize the sludge. Depositary Authority as established under the Budapest Intake air passes in through Small holes located around the Treaty according to 37 CFR 1.803(a)(1), such as the Ameri central mouth piece. This air flow creates an upward current can Type Culture Collection (ATCC) in Manassas, Va., and that allows the essential oil vapor to be inhaled. the Agricultural Research Service Culture Collection 0585. The vaporizer may also comprise a selector switch (NRRL, acronym based on former name) in Peoria, Illinois, which is designed to select the desired cannabis dosage. This or a depository recognized to be suitable by the United States Switch communicates with the microchip to control how Patent and Trademark Office under 37 CFR 1.803(a)(2). many of the activation sites on the dosing strip are activated 0593. To satisfy the enablement requirements of 35 U.S.C. and activates the activation sites in any possible combination. 112, and to certify that the deposit of the isolated strains (i.e., In non-limiting examples, the Switch activates each of the cannabis varieties) of the present invention meets the criteria four sites individually, one, two, three or four of the sites set forth in 37 CFR 1.801-1809 and Manual of Patent Exam consecutively or serially, or one, two, three, or four sites with ining Procedure (MPEP) 2402-2411.05, Applicants hereby a delay between them. The orders in which the activation sites make the following statements regarding the deposited can are activated, and/or the delay between the activation of one nabis varieties: or more sites, are calculated based on dosage studies. 0594. If the deposit is made under the terms of the Budap est Treaty, the instant invention will be irrevocably and with Example 14 out restriction released to the public upon the granting of a patent. Zero-Point Delivery Doses 0595. If the deposit is made not under the terms of the 0586. The vaporization device described in the above Budapest Treaty, Applicant(s) provides assurance of compli example is designed to work with dosage Strips engineered ance by following statements: specifically for efficient vaporization at the particular voltage 0596 1. During the pendency of this application, access to and current Supplied by the device. An example of a dosing the invention will be afforded to the Commissioner upon strip is shown in FIG. 6. request; 0587. The strips are composed of a non-conductive mate 0597 2. All restrictions on availability to the public will be rial Such as ceramic or glass, and contain sludge from a whole irrevocably removed upon granting of the patent under con plant liquid-gas extract at the particular resistance that is ditions specified in 37 CFR 1.808: vaporized by the device. The dosage strips comprise an alu 0598. 3. The deposit will be maintained in a public reposi minum conductor with four (4) or more resistance sites, each tory for a period of 30 years or 5 years after the last request or of which consists of bundles of frayed aluminum which con for the effective life of the patent, whichever is longer; US 2014/029851 1 A1 Oct. 2, 2014 50
0599 4. A test of the viability of the biological material at 17. The cannabis plant part of claim 16, wherein said plant the time of deposit will be conducted by the public depository part is selected from the group consisting of seeds, trichomes, under 37 CFR 1.807; and Sun leaves, and inflorescences. 0600 5. The deposit will be replaced if it should ever 18. The cannabis plant, plant part, tissue, or cell of claim become unavailable. 16, wherein the cannabis plant, plant part, tissue, or cell 0601 Access to this deposit will be available during the comprises a relative alpha pinene content that is greater than pendency of this application to persons determined by the about 16% of the total content of detected terpenes. Commissioner of Patents and Trademarks to be entitled 19. The cannabis plant, plant part, tissue, or cell of claim thereto under 37 C.F.R. S.1.14 and 35 U.S.C. S 122. Upon 16, wherein the cannabis plant, plant part, tissue, or cell granting of any claims in this application, all restrictions on comprises a relative linalool content that is greater than about the availability to the public of the variety will be irrevocably 14.5% of the total content of detected terpenes. removed by affording access to a deposit of at least 2,500 20. The cannabis plant, plant part, tissue, or cell of claim seeds of the same variety with the depository. 16, wherein the cannabis plant, plant part, tissue, or cell 0602 Unless defined otherwise, all technical and scien comprises a relative beta-caryophyllene content that is tific terms herein have the same meaning as commonly under greater than about 31% of the total content of detected terpe stood by one of ordinary skill in the art to which this invention CS. belongs. Although any methods and materials, similar or 21. The cannabis plant, plant part, tissue, or cell of claim equivalent to those described herein, can be used in the prac 16, wherein the cannabis plant, plant part, tissue, or cell tice or testing of the present invention, the non-limiting exem comprises a relative terpinolene content that is greater than plary methods and materials are described herein. about 12.5% of the total content of detected terpenes. 0603 All publications and patent applications mentioned 22. A cannabis plant, plant part, tissue, or cell comprising: in the specification are indicative of the level of those skilled a) a cannabidiol (CBD) content that is greater than 1.5% by in the art to which this invention pertains. All publications and weight; patent applications are herein incorporated by reference to the b) a tetrahydrocannabinol (THC) content that is greater same extent as if each individual publication or patent appli than 2.0% by weight; and cation was specifically and individually indicated to be incor c) a myrcene terpene content less than about 0.16% by porated by reference. Nothing herein is to be construed as an weight; admission that the present invention is not entitled to antedate wherein the cannabinoid and terpene content is calculated such publication by virtue of prior invention. based on dry weight of the inflorescence. 0604 Many modifications and other embodiments of the 23. The cannabis plant part of claim 22, wherein said plant inventions set forth herein will come to mind to one skilled in part is selected from the group consisting of seeds, tri the art to which these inventions pertain having the benefit of chomes, Sun leaves, and inflorescences. the teachings presented in the foregoing descriptions and the 24. The cannabis plant, plant part, tissue, or cell of claim associated drawings. Therefore, it is to be understood that the 22, wherein the cannabis plant, plant part, tissue, or cell inventions are not to be limited to the specific embodiments comprises an alpha pinene terpene content that is greater than disclosed and that modifications and other embodiments are about 0.17% by dry weight. intended to be included within the scope of the appended 25. The cannabis plant, plant part, tissue, or cell of claim claims. Although specific terms are employed herein, they are 22, wherein the cannabis plant, plant part, tissue, or cell used in a generic and descriptive sense only and not for comprises a linalool terpene content that is greater than about purposes of limitation. 0.07% by dry weight. 0605 While the invention has been described in connec 26. The cannabis plant, plant part, tissue, or cell of claim tion with specific embodiments thereof, it will be understood 22, wherein the cannabis plant, plant part, tissue, or cell that it is capable of further modifications and this application comprises a beta-caryophyllene terpene content that is is intended to cover any variations, uses, or adaptations of the greater than about 0.15% by dry weight. invention following, in general, the principles of the invention 27. The cannabis plant, plant part, tissue, or cell of claim and including Such departures from the present disclosure as 22, wherein the cannabis plant, plant part, tissue, or cell come within known or customary practice within the art to comprises a terpinolene terpene content that is greater than which the invention pertains and as may be applied to the about 0.07% by dry weight. essential features hereinbefore set forth and as follows in the 28. A method of breeding a cannabis plant, comprising a Scope of the appended claims. cannabidiol (CBD) content that is greater than 1.5% by weight, a tetrahydrocannabinol (THC) content that is greater 1.-15. (canceled) than 2.0% by weight, and a relative myrcene content of less 16. A cannabis plant, plant part, tissue, or cell comprising: than about 31% of the total content of detected terpenes, said a) a cannabidiol (CBD) content that is greater than 1.5% by method comprising: weight; (i) making a cross between a first cannabis plant and a b) a tetrahydrocannabinol (THC) content that is greater second cannabis plant to produce an F1 plant, wherein than 2.0% by weight; and the first cannabis plant is the cannabis plant of claim 16; c) a relative myrcene content of less than about 31% of the (ii) harvesting the resulting seed; total content of detected terpenes; (iii) growing said seed; and wherein said detected terpenes include terpinolene, myrcene, (iv) selecting for the desired phenotypes; limonene, linalool, caryophyllene oxide, beta-caryophyllene, wherein the resulting selected cannabis plant is a cannabis and alpha pinene; and wherein the content of each cannab plant comprising a cannabidiol (CBD) content that is greater inoid and terpene is calculated based on dry weight of the than 1.5% by weight, a tetrahydrocannabinol (THC) content inflorescence. that is greater than 2.0% by weight, and a relative myrcene US 2014/029851 1 A1 Oct. 2, 2014
content of less than about 31% of the total content of detected (iii) allowing said cannabis seed, cutting, or plant to pro terpenes, wherein said detected terpenes include terpinolene, duce a cannabis plant; myrcene, limonene, linalool, caryophyllene oxide, beta wherein the resulting selected cannabis plant is a cannabis caryophyllene, and alpha pinene; and wherein the content of plant comprising a cannabidiol (CBD) content that is greater each cannabinoid and terpene is calculated based on dry than 1.5% by weight, a tetrahydrocannabinol (THC) content weight of the inflorescence. that is greater than 2.0% by weight, and a relative myrcene 29. A cannabis plant, plant pan, or cell, produced by the content of less than about 31% of the total content of detected methods of claim 28. terpenes, wherein said detected terpenes include terpinolene, 30. A method of producing a cannabis plant, comprising a myrcene, limonene, linalool, caryophyllene oxide, beta cannabidiol (CBD) content that is greater than 1.5% by caryophyllene, and alpha pinene; and wherein the content of weight, a tetrahydrocannabinol (THC) content that is greater each cannabinoid and terpene is calculated based on dry than 2.0% by weight, and a relative myrcene content of less weight of the inflorescence. than about 31% of the total content of detected terpenes, said 31. A cannabis plant, plant part, or cell, produced by the method comprising: methods of claim 28. (i) obtaining a cannabis seed, cutting, or plant cell capable 32. The cannabis plant, or an asexual clone of said can of growing into a cannabis plant from the plant or plant nabis plant, or a plant part, tissue, or cell thereof of claim 16. part of claim 16; 33. The cannabis plant, or an asexual clone of said can (ii) placing said cannabis seed, cutting, or plant cell in an nabis plant, or a plant part, tissue, or cell thereof of claim 22. environment conducive to plant growth; k k k k k