Compounds and methods for the prevention of melanoma

Abstract

Disclosed herein are pan-RAF inhibitors, pharmaceutical compositions comprising such compounds, and methods of using such compounds for the inhibition and / or prevention of melanoma, including for example, primary and locally recurrent melanoma.

Description

COMPOUNDS AND METHODS FOR THE PREVENTION OF MELANOMACROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority under 35 U. S. C. § 119 to U. S. Provisional Patent Application No. 63 / 752,614 filed January 31, 2025, which is hereby incorporated by reference in its entirety.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0002] This invention was made with government support under grant / contract number R01 CA287193 awarded by the National Institutes of Health. The government has certain rights in the invention.FIELD

[0003] Disclosed herein are compounds, pharmaceutical compositions and methods for the inhibition and / or prevention of melanoma, including for example, primary and locally recurrent melanoma.BACKGROUND

[0004] Melanoma constitutes only a small percentage (about 4%) of dermatologic cancers, but it is responsible for a significant percentage (about 80%) of deaths.

[0005] Sun avoidance and protection practices are also encouraged for melanoma prevention. However, no methods exist to reverse the harm caused by previous UV exposures. This is especially relevant given the significant melanoma risk attributed to childhood and adolescent sunburn.

[0006] Early detection plays a critical role in the survival of individual s with cutaneous melanoma. The five-year survival rate for patients diagnosed with advanced disease is considerably lower than for those with early-stage lesions (32 vs, >99%, respecti vely). Melanoma 5-year survival rates depend on the stage of diagnosis (local melanoma (>95%), regional melanoma (75%) and metastatic (32%). Nevertheless, melanoma detection poses many challenges, particularly for individuals with numerous nevi. Metastatic melanoma can also arise in the absence of an identified primary lesion. Identifying early-stage melanoma is clinicallychallenging. Missed melanoma diagnoses are the most common malpractice claim in pathology. Inter-pathologist diagnostic consensus is only ~83% for biopsied moles. Only 1 melanoma case is identified out of every 14 to 53 skin biopsies. Two-thirds of melanomas arise in the absence of a prior mole. Finally, up to 20% of early stage (stage II) melanomas recur, even after surgical removal. There remains a need for novel approaches to inhibit and / or prevent melanoma at the earliest stages.SUMMARY

[0007] Disclosed herein are compounds and pharmaceutical compositions useful in the inhibition and / or prevention of melanoma. Also disclosed are methods of inhibiting and / or preventing melanoma comprising administering a pan-RAF inhibitor.

[0008] In some embodiments, the melanoma is an NRAS or BRAF mutant melanoma. In some embodiments, the melanoma is a localized primary and / or locally recurrent melanoma.

[0009] In one aspect, disclosed herein is a method of reducing the size of a melanoma precursor lesion in a subject in need thereof, the method comprising: non-systemically administering a pan-RAF inhibitor to the melanoma precursor lesion; wherein the method does not comprise co-administering a downstream ATP competitive inhibitor of the MAP kinase pathway; and wherein the melanoma precursor lesion has not been diagnosed as melanoma.

[0010] In another aspect, disclosed herein is a method of inhibiting progression of a melanoma precursor lesion to melanoma in a subject in need thereof, the method comprising: non-systemically administering a pan-RAF inhibitor to the melanoma precursor lesion; wherein the method does not comprise co-administering a downstream ATP competitive inhibitor of the MAP kinase pathway; and wherein the melanoma precursor lesion has not been diagnosed as melanoma.

[0011] In yet another aspect, disclosed herein is a method of preventing melanoma in a subject in need thereof, the method comprising: non-systemically administering a pan-RAF inhibitor to the subject; wherein the method does not comprise co-administering a downstream ATP competitive inhibitor of the MAP kinase pathway; and wherein the subject does not have a current melanoma diagnosis,

[0012] In another aspect, disclosed herein is a method for inhibiting recurrence of melanoma in a border region remaining after excision of a local melanoma in a subject, the methodcomprising: non-systemically administering a pan-RAF inhibitor to the border region of the subject following the excision of the focal melanoma; wherein the method does not comprise coadministering a downstream ATP competitive inhibitor of the MAP kinase pathway.

[0013] In one embodiment of any aspect disclosed herein, the pan RAF inhibitor is AZ628, avutometinib, belvarafenib, brimarafenib, CCT196969, CCT241161, exarafenib, GNE-9815, IHMT-RAF-128, JZP81538, lifirafenib, LSN3074753, naporafenib, pan-RAF kinase inhibitor 1, RAF709, SJ-C1044, TBAP-001, tovorafenib, TAK-632 or a therapeutically effective derivative or analog thereof.BRIEF DESCRIPTION OF THE FIGURES

[0014] FIGS 1A, IB, 1C, and I D show that TN and TB mouse models mirror the genetic and environmental etiologies of human nevi and melanoma FIG 1 A shows the TN mouse model is homozygous for a melanocyte-specific, tamoxifen-inducible CRE transgene (Tyr:: CreER(T2)) and a conditional NrasQ61Ror Nras61Hknock-in allele LSL-NrasQ6‘kH)'. FIG. 1B shows TB mice carry a single BrafV637Econditional allele (LSL-BrafCA; 50) and are homozygous for a conditional p16INK4aknockout allele (p16L; 51) and Tyr:: CreER(T2). Note that BrafV637Eis the murine equivalent of human BRAFV600E. Open triangles represent Lox P sites and the V637E mutation is indicated by a star. The Kaplan-Meier curve shows that a single UV exposure accelerates TB melanoma onset by 60%. ** p = 0.01, log-rank test. FIG. 1C shows the treatment and UV irradiation scheme for the mice FIG. ID are representative images of spontaneous nevi in TN and TB mice. FIGS. 1E and 1F are Kaplan-Meier curves showing that a single UV exposure accelerates melanoma onset in the TN and TB models. * p<0.05, ** p = 0.01, f p<0.0001, log¬ rank test, UV vs. No UV for each genotype.

[0015] FIG. 2 is a scheme illustrating how type II RAF inhibitors (pan-RAF inhibitors) limit paradoxical MAPK activation.

[0016] FIGS. 3A and 3B show that the melanomagenic potential of NRAS mutants correlates with BRAF binding affinity. FIG. 3 A is a schematic of a BRET assay. FIG. 3B is a graph of BRET50for different Q61 mutations (R = arginine, K = lysine, L = leucine, H = histidine, and P = proline).

[0017] FIGS. 3C-3D show representative immunoblots (FIG. 3C) and graphical summary of densitometry measurements (FIG. 3D) of phosphorylated ERK (pERK) as a fraction of totalERK in lysates from human melanoma cell lines treated for 1 hour with vehicle (V, DMSO) or a pan-RAF inhibitor: (belvarafenib (B), exarafenib (E), or naporafenib (N)).

[0018] FIG. 4A shows an experimental design used to assess the impact of transient pan- RAF inhibition on NRAS mutant nevi using 6-week-old TN61Hmice.

[0019] FIG. 4B shows dorsal photographs of depilated TN61Hmice 1 week after completing treatment with vehicle or exarafenib as described in FIG. 4A. Black color indicates pigmentation from melanocytes.

[0020] FIG. 4C is a box and whisker plot of the number of melanocytes per area in Fontana- Masson-stained dorsal skin sections in tissues harvested from the mice of FIG. 4B. Fontana-Masson stains melanin-containing cells. Dots represent biological replicates, t-test ** p < 0.01 student’s t-test

[0021] FIG. 4D shows representative images of the footpad of TN61Hmice five weeks after completing treatment with vehicle, exarafenib, or naporafenib as described in FIG. 4A.

[0022] FIG. 4E shows a graph of footpad pigmentation as scored by at least four treatment- blinded reviewers. The average reviewer score for each mouse is indicated by a dot and the mean is shown as a line.

[0023] FIG. 5 A shows an experimental design to determine how transient pan-RAF inhibition impacts NRAS mutant melanoma formation.

[0024] FIGS. 5B displays the Kaplan-Meier curves of tumor formation in TN6’11mice treated as described in FIG. 5A.

[0025] FIG. 5C displays the Kaplan-Meier curves of tumor formation in TN61Rmice treated as described in FIG. 5 A.

[0026] FIG. 6A is a schematic of the experiment showing TN61Rmice were treated with 4-hydroxytamoxifen (4OHT) to induce melanocyte-specific oncogenic NRAS expression on postnatal days 1 and 2. They were then subjected to a non-burning dose of UV on post-natal day 3. At 3 weeks of age, the tails of the mice were treated topically with vehicle (W06), 1% exarafenib, 1% naporafenib, or 1% avutometinib and harvested for immunoblotting 1 hour or 3 hours later.

[0027] FIG. 6B shows representative immunoblots using the protein from the treated mouse tails harvested at 1 hour.

[0028] FIG. 6C shows quantification of MAPK pathway activity, phosphorylated ERK / total ERK, as measured from the immunoblots in FIG, 6B. Each dot represents a single mouse. Bars shown mean ± SEM..

[0029] FIG. 6D shows representative immunoblots using the protein from the treated mouse tails harvested at 3 hours.

[0030] FIG. 6E shows quantification of MAPK pathway target, DUSP6, normalized to a-Tubulin as measured from the immunoblots shown in FIG. 6D. Each dot represents a single mouse. Bars shown mean ± SEM.

[0031] FIG. 6F shows Kaplan-Meier curves comparing topical treatment for five consecutive days with vehicle (W06), 0.5-1% exarafenib, or 1-3% dabrafenib.

[0032] FIGS. 7A-B show that topical pan-RAF inhibition slows the growth of established NRAS mutant melanomas. TN^ melanomas (7-8 mm in diameter) were treated topically with vehicle (n = 8), exarafenib (n = 7) or avutometinib (n = 5) once daily for up to 10 days. Tumor size was measured daily using digital calipers. FIG. 7A is a graph of change in tumor size over days of treatment. Data show mean percent change in tumor volume from baseline ± SEM. FIG. 7B is a waterfall plot showing percent change in tumor size from baseline to the animal endpoint for the tumors in FIG. 7A.DETAILED DESCRIPTION

[0033] Embodiments described herein can be understood more readily by reference to the following detailed description and examples and their previous and following descriptions. Elements and methods described herein, however, are not limited to the specific embodiments presented in the detailed description and examples. It should be recognized that these embodiments are merely illustrative of the principles of the present invention. Numerous modifications and adaptations will be readily apparent to those of skill in the art without departing from the spirit and scope of the invention.

[0034] In addition, all ranges disclosed herein are to be understood to encompass any and all subranges subsumed therein. For example, a stated range of "1.0 to 10.0" should be considered to include any and all subranges beginning with a minimum value of 1 0 or more and ending with a maximum value of 10.0 or less, e g., 1.0 to 5.3, or 4.7 to 10.0, or 3.6 to 7.9.

[0035] When a range of integers is given, the range includes any number falling within the range and the numbers defining ends of the range. For example, when the terms "integer from 1 to 20" is used, the integers included in the range are 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc., up to and including 20. All ranges disclosed herein are also to be considered to include the end points of the range, unless expressly stated otherwise. For example, a range of "between 5 and 10" should generally be considered to include the end points 5 and 10.

[0036] Further, when the phrase "up to" is used in connection with an amount or quantity, it is to be understood that the amount is at least a detectable amount or quantity. For example, a material present in an amount "up to" a specified amount can be present from a detectable amount and up to and including the specified amount.

[0037] Furthermore, the terms "substantially," "approximately," and "about," as used herein when referring to a measurable value such as an amount of a compound or agent of this invention, dose, time, temperature, and the like, is meant to encompass variations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of the specified amount. The term "consists essentially of' (and grammatical variants) shall be given its ordinary' meaning and shall also mean that the composition or method referred to can contain additional components as long as the additional components do not materially alter the composition or method. The term "consists of (and grammatical variants) shall be given its ordinary' meaning and shall also mean that the composition or method referred to is closed to additional components. The term "comprising" (and grammatical variants) shall be given its ordinary meaning and shall also mean that the composition or method referred to is open to contain additional components.

[0038] It is also to be understood that the article "a" or "an" refers to "at least one," unless the context of a particular use requires otherwise.

[0039] Also as used herein, "and / or" refers broadly to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative ("or").

[0040] Compounds, pharmaceutical compositions including the compounds, and methods of preparation and uses thereof are disclosed. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary' skill in the art to which this subject matter belongs. The terminology used in the description of the subject matter herein is for the purpose of describing particular embodiments only and is notintended to be limiting of the subject matter. The present disclosure will be better understood with reference to the following definitions.Definitions

[0041] “Amino acid” or “amino acid residue” as used herein typically refers to a naturally- occurring amino acid. The one letter code is used herein to refer to the respective amino acid. As used herein, a “charged amino acid” is an amino acid that is negatively or positively charged. “Negatively charged amino acids” are aspartic acid (D) and glutamic acid (E).“Positively charged amino acids” are arginine (R) lysine (K) and histidine (H). “Polar amino acids” are all amino acids that form hydrogen bonds as donors or acceptors. These are all charged amino acids and asparagine (N), glutamine (Q), serine (S), threonine (T), tyrosine (Y) and cysteine (C). “Pol r uncharged amino acids” are asparagine (N), glutamine (Q), serine (S), threonine (T), tyrosine (Y) and cysteine (C). “Amphipathic amino acids” are tryptophan (W), tyrosine (Y) and methionine (M). “Aromatic amino acids” are phenylalanine (F), tyrosine (Y), and tryptophan (W). “Hydrophobic amino acids” are glycine (G), alanine (A), valine (V), leucine (L), isoleucine (I), proline (P), phenylalanine (F), methionine (M) and cysteine. “Small amino acids” are glycine (G), alanine (A), serine (S), proline (P), threonine (T), aspartic acid (D) and asparagine (N). In some embodiments, the NRAS and / or BRAF mutations described herein involve a point mutation affecting one or more amino acids.

[0042] “Analog” as used herein refers to a compound having a similar structure and / or mechanism of action to the reference drug but differing in at least one structural element.

[0043] “Apoptosis” as used herein refers to a regulated process of cell death wherein a dying cell displays a set of wel I -characterized biochemical hallmarks that can include cell membrane blebbing, cell soma shrinkage, chromatin condensation, and DNA laddering, as well as any caspase- mediated cell death.

[0044] “Combination therapy” or “co-administration” as used herein refers to a method of treatment comprising administering to a subject at least two therapeutic agents, optionally as one or more pharmaceutical compositions. For example, a combination therapy may comprise the administration of a single pharmaceutical composition comprising at least two therapeutic agents and one or more pharmaceutically acceptable carriers, excipients, diluents, and / or surfactants. A combination therapy may comprise the administration of two or more pharmaceuticalcompositions, each composition comprising one or more therapeutic agents and one or more pharmaceutically acceptable carriers, excipients, diluents, and / or surfactants.[00451 “Dysplastic nevus” as used herein refers to an atypical mole or a mole whose appearance is different from that of common moles Dysplastic nevi are generally larger than ordinary moles or have irregular and indistinct borders. Their color frequently is often not uniform and ranges from pink to dark brown; they usually are flat, but parts may be raised above the skin surface.

[0046] "‘Excipient” as used herein refers to any pharmaceutically acceptable additive, carrier, diluent, adjuvant, or other ingredient, other than the active pharmaceutical ingredient (API), which is typically included for formulation and / or administration to a subject.

[0047] “Melanocytes” are pigment cells normally present in the epidermis, in adnexal structures including hair follicles, and sometimes in the dermis, as well as extracutaneous sites such as the mucosa, meninx, conjunctiva, and uvea.

[0048] “Melanoma” as used herein refers to a cancer characterized by the uncontrolled growth of melanocytes. The melanoma may be any form of melanoma, such as superficial spreading melanoma, nodular melanoma or acral lentiginous melanoma. Cancerous melanocytes can also be referred to as a “melanoma lesion.”

[0049] Melanoma precursor lesion” refers to an accumulation of melanocytes that can undergo development that is benign, locally aggressive or malignant. Melanoma precursor lesion can encompass both benign cells, such as nevi, and malignant cells, melanoma. In some embodiments, a melanoma precursor lesion comprises an NR. AS- and / or BRAF -mutant lesion predisposed to melanoma.

[0050] “ elanomagenesis” as used herein refers to the transformation of normal melanocytes into malignant cells. In some embodiments, a method is disclosed for eliminating melanomagenesis. In some preferred embodiments, a method is disclosed for reducing, inhibiting or preventing melanomagenesis.

[0051] MEK” as used herein refers to mitogen-activated protein kinase / extracellular signal- regulated kinase.

[0052] “MEK inhibitor” or “MEK antagonist” as used herein refers to a compound that binds to and / or inhibits MEK, but not RAF, with measurable affinity. In one embodiment, a MEK inhibitor is an ATP-competitive MEK inhibitor. ATP-competitive MEK inhibitors include butare not limited to BI-847325 (Boehringer Ingelheim), PD 184352 (CT-1040, Pfizer), PD 0325901 (Pfizer), PD 098059 (2-(2'-amino-3'-methoxphenyl)- oxanaphthal en-4-one) (Pfizer), and U0126 (l,4-diamino-2,3-dicyano-l,4-bis(2- aminophenylthio)butadiene, Sigma). In one embodiment, a MEK inhibitor is an allosteric, non-ATP competitive inhibitor of MEK which does not simultaneously bind RAF Allosteric MEK inhibitors include but are not limited to ARRY-142886 (AZD6244), binimetinib (ARRY-438162), cobimetinib (GDC-0973), pimasertib, RDEA119, R04987655, refametinib, selumetinib (AZD6244), and trametinib. In some preferred embodiments, the method disclosed herein excludes co-administration of a MEK inhibitor. In one embodiment, a method disclosed herein excludes co-administration of an ATP-competitive MEK inhibitor. In another embodiment, a method disclosed herein excludes co-administration of an allosteric non-ATP competitive MEK inhibitor that does not simultaneously bind RAF. In one embodiment, the methods disclosed herein exclude co-administration of one or more of PD184352 (CI-1040), RO-4987655, GDC-0623, TAK-733, AZD-8330, AS-701255, AS-701173, BI 847325, PD 184352, PD 0325901, PD 098059, U0126, ARRY-142886, WX-554, CH5126766 (R05126766), G-573, Arry 300, SHR 7390, MSC2015103B (AS-703988), LY 2228820 (Ralimetinib), and CS 3006 binimetinib, cobimetinib, pimasertib, RDEA119, R04987655, refametinib, selumetinib, and trametinib.

[0053] Mutation” as used herein refers to a substitution, deletion, and insertion of one or more nucleotides encoding a polynucleotide. For example, up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 40, 50, or more nucleotides in a polynucleotide (cDNA, gene) can be substituted, deleted, and / or inserted. A mutation can affect the coding sequence of a gene or its regulatory sequence. A mutation can also affect the structure of the genomic sequence or the structure / stabilitv of the encoded mRNA.

[0054] “Nevus” (plural nevi) is used herein to refer to a mole, which is a benign growth on the skin formed by clusters of mel anocytes, the cells that produce pigment. A skin lesion suspected of being melanoma may be a mole or nevus. Melanoma lesions are usually, but not always, larger than 6 mm in diameter. Not all melanoma lesions are visually detectable.

[0055] " NRAS” as used herein refers to neuroblastoma RAS viral oncogene homolog, a membrane-bound RAS small G protein family member that initiates activation of the RAF family of serine / threonine kinases, including BRAF, through RAF recruitment to the plasma membrane, thereby triggering sequential downstream activation of the protein kinases MEK andthen ERK I and ERK2, as well as Myc, a transcription factor activated by MEK-> ERK-> Myc. NRAS mutations are found in about 20% of all melanoma cases.[00561 “Prevention” as used herein refers to partially or completely delaying the onset of an infection, disease, disorder and / or condition; partially or completely delaying the onset of one or more symptoms, features, or clinical manifestations of a particular infection, disease, disorder, and / or condition partially or completely delaying progression from an infection, a particular disease, disorder and / or condition; and / or decreasing the risk of developing pathology associated with an infection, disease, disorder, and / or condition.

[0057] “RAF” as used herein refers to Rapidly Accelerated Fibrosarcoma proteins RAF molecules play a critical role in cell signaling through their integral impact on the RAS / RAF / MEK / ERK signaling pathway,

[0058] “RAF -inhibitor” as used herein refers to molecules that prevent RAF signaling through the MEK / ERK pathway. “Pan-RAF inhibitors” as used herein refers to a class of RAF inhibitors that target multiple RAF isoforms (e g. BRAF, CRAF, and ARAF or any mutations thereof) simultaneously. In one embodiment, pan-RAF inhibitor as used herein includes compounds that inhibit both protomers in a RAF dimer. In one embodiment, pan-RAF inhibitor as used herein includes a RAF / MEK inhibitor, which binds to both RAF and MEK proteins. The methods disclosed herein comprise the use of at least one pan-RAF inhibitor.

[0059] “Side effects” as used herein refers to unintended, and undesirable, consequences arising from active agent therapy In one embodiment, the methods disclosed herein result in minimal or no systemic side effects. In one embodiment, the methods disclosed herein result in reduced side effects compared to standard of care treatment.

[0060] “SHP2” or “Shp2” as used herein refers to a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene that contributes to multiple cellular functions including proliferation, differentiation, cell cycle maintenance and migration. SHP2 is involved in signaling through the RAS-mitogen-activated protein kinase (MAPK), the JAK-STAT and / or the phosphoinositol 3- kinase- AKT pathways. In one embodiment, at least one additional therapeutic agent administered according to the method disclosed herein is a Shp2 inhibitor such as, for example, an upstream Shp2 inhibitor. In one embodiment, the present method excludes co-administration of a Shp2 inhibitor.

[0061] “Synergistically effective” or “synergistic effect” indicates that two or morecompounds that are therapeutically effective (e.g., compounds of Formula (T)-(IV) and at least one additional therapeutic agent), and when used in combination, provide improved therapeutic effects greater than the additive effect that would be expected based on the effect of each compound used by itself.

[0062] “Subject” as used herein refers to animals such as mammals, e.g., humans, apes, monkeys, dogs, cows, horses, pigs, sheep, goats, cats, mice, rabbits, rats, and transgenic non¬ human animals. In one embodiment, a subject “in need of’ the compounds, compositions or methods disclosed herein is a subject that has been identified as needing or benefiting from administration of the compounds or compositions disclosed herein. In one embodiment, a subject in need is experiencing a disease state and / or is a subject anticipated to experience a disease state. Thus, the compounds, compositions and methods disclosed herein can be used for prophylactic as well as therapeutic uses. In one embodiment, the subject is a human, e g., a human suffering from, at risk of suffering from, or potentially capable of suffering from cancer, such as melanoma, including for example an NRAS- and / or BRAF mutant melanoma.

[0063] “Therapeutically effective amount" as used herein means an amount of a compound, or combination of compounds, that ameliorates, attenuates or eliminates one or more symptoms of a particular disease or condition, or that delays or reduces the progression of a disease or condition or that prevents or delays the onset of one or more symptoms of a particular disease or condition.

[0064] As used herein, “delaying progression of melanoma” means to defer, hinder, slow, retard, stabilize, and / or postpone the development of melanoma. A delay can be of varying lengths of time, depending on the history' of the disease and / or individual being treated. A sufficient or significant delay can, in effect, encompass prevention, in that the individual does not develop the disease. In one embodiment, local recurrence of melanoma is delayed. In another embodiment, the development of metastasis is delayed. In one embodiment, progression is delayed when the size of a melanoma precursor lesion or a melanoma lesion has not grown larger or has shrunk based on a comparison of an initial measurement of the sum of the longest diameters (SLD) of the identified lesion (the baseline SLD) and the SLD measured after non-systemic administration of the pan-RAF inhibitor as described herein.

[0065] According to a general model, the first step in melanoma development is represented by the clustering of melanocytes, leading to the formation of a benign nevus. Thedevelopment of cytological atypia within a benign nevus results in the formation of dysplastic nevi (second step). A dysplastic nevus can either remain unchanged, regress or develop into a radial growth phase (RGP, third step) melanoma; this last one can then progress into a more aggressive vertical growth phase (VGP, fourth step) melanoma. Failure to progress through one or more of these steps in one embodiment, is an outcome of the method of prevention outlined herein. For example, in one embodiment, non-systemic administration of a pan-RAF inhibitor according to the methods disclosed herein is responsible for the failure to progress from step 1 to step 2, from step 2 to step 3, and / or from step 3 to step 4.

[0066] As used herein “non-systemic administration” or “non-systemically administered” refers to administration of an active agent or composition comprising an active agent to a subject, such that the active agent does not significantly enter the subject’s blood stream. An active agent that is administered non-systemically can be found generally in therapeutic concentrations in the local region of administration. In contrast, systemic administration of a compound refers to administration such that the compound significantly enters the subject’s blood stream. Systemic administration can include, but is not limited, to oral, intravenous, intraperitoneal and intramuscular administration.

[0067] As used herein, “topical” refers to application of a suitable compound (e.g. active agent) or composition comprising an active agent to the skin In one embodiment, “topical” means application of an active agent or composition comprising an active agent to the skin with adequate penetration of the epidermis or dermis to treat or prevent a lesion or a disease of the epidermis and / or dermis. In one embodiment, the topically administered compound or composition penetrates the epidermis or dermis without significant systemic exposure. In such embodiments, there is no intent to treat or prevent a disease via systemic therapeutic dosing nor to treat or prevent a disease of another organ system.

[0068] Melanoma is generally characterized by Stages: In Stage 0 melanoma, the malignant tumor is confined to the epidermis and has not grown into the second layer of skin, the dermis. Stage 0 melanoma is not considered invasive melanoma and is sometimes referred to as “ / « situ” melanoma. In Stage I melanoma, the cancer cells are in both the epidermis and the dermis, characterized as invasive melanoma. Stage I melanoma is still considered local melanoma, meaning it has not spread beyond the primary tumor. A melanoma tumor is considered Stage I if it is up to 2 mm thick with or without ulceration. Tumor thickness (also called Breslow Depth,measured in millimeters) refers to how deeply the tumor has penetrated the skin. Tumors that are less than 0.8 mm without ulceration are Stage IA. Tumors that are greater than 1.0 mm and up to 2.0 mm without ulceration are Stage IB. In Stage II melanoma, the cancer cells are in both the epidermis and the dermis, that is invasive melanoma, but there is no evidence the cancer has spread to lymph nodes or distant sites (metastasis). Stage II melanoma is local melanoma, meaning it has not spread beyond the primary tumor. There are three subgroups of Stage II melanoma: IIA, IIB, and IIC. In Stage IIA, the tumor is 1.01 -• 2.0 mm thick with ulceration, or tumor is 2.01 - 4.0 mm thick without ulceration. In Stage IIB, the tumor is 2.01 - 4.0 mm thick with ulceration, or tumor is greater than 4.0 mm thick without ulceration In Stage IIB, the tumor is greater than 4.0 mm thick with ulceration. In Stage III, main tumor can be any thickness, and it might or might not be ulcerated; The cancer has spread to nearby lymph nodes and / or it has spread to very small areas of nearby skin (satellite tumors) or to skin lymphatic channels around the tumor. In Stage III, there are no signs the cancer has spread to distant parts of the body. In Stage IV, the main tumor can be any thickness, and it might or might not be ulcerated; the cancer might or might not have spread to nearby lymph nodes The cancer has spread to distant parts of the body, such as: (a) areas of skin or lymph nodes in other parts of the body; (b) lung(s); (c) any other organs outside the central nervous system; (d) the central nervous system, including the brain, spinal cord, and the coverings of the brain and spinal cord.

[0069] The presently disclosed subject matter will now be described more fully hereinafter. However, many modifications and other embodiments of the presently disclosed subject matter set forth herein will come to mind to one skilled in the art to which the presently disclosed subject matter pertains having the benefit of the teachings presented in the foregoing descriptions. Therefore, it is to be understood that the presently disclosed subject matter is not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. In other words, the subject matter described herein covers all alternatives, modifications, and equivalents. In the event that one or more of the incorporated literature, patents, and similar materials differs from or contradicts this application, including but not limited to defined terms, term usage, described techniques, or the like this application controls. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinaryskill in this field All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.I. General

[0070] Most nevi (moles) contain an oncogenic NRAS or BRAF mutation, but never progress to form cancer. The nevus-to-melanoma transition requires enhanced RAF engagement and signaling. MAPK> ERK signaling increases in human nevi that progress to form melanoma and NRAS mutants that induce greater RAF activation are more melanomagenic in mice.

[0071] While the methods disclosed herein apply to all subjects at risk of developing melanoma, three well-defined groups are particularly notable:

[0072] 1. Children born with large congenital nevi (LCN): These children, who make up approximately 1 in 20,000 births, have a 4-10% increased risk for melanoma development and often undergo multiple surgeries to remove and graft non-melanocytic skin in place of these lesions8’9. Notably, 57-95% of LCN contain oncogenic NRAS mutations, and their atypical appearance makes it difficult to distinguish benign growth from malignancy10.

[0073] 2. Individuals with atypical or multiple nevus syndromes: These individuals have a 10-to 500-fold increased risk of melanoma11and face challenges with monitoring due to the number and atypical features of their nevi.

[0074] 3. Early-stage melanoma survivors: Thousands of Americans are diagnosed with early-stage melanoma each year12. Such diagnoses include Stages 0 and I and can sometimes include Stage II. Although primarily treated with surgical excision, up to 20% of these tumors recur.7

[0075] Data from both patient and animal models show that some cutaneous nevi are sensitive to MARK inhibition. For example, MEK inhibitors block the paradoxical MARK activation caused by type I BRAF inhibitors. This leads to hypopigmentation and the regression of melanocytic nevi20,25,26. Multiple studies of mouse and human NRAS-mutant giant congenital nevi also find that MAPK inhibition induces melanocytic cell death at early developmental stages27'29. Without being bound by theory, these data support the idea that inhibiting MAPK signaling in BRAF- and / or NRAS-mutant nevi may eliminate cancer progenitors from the skin.

[0076] The three RAF homologs, ARAF, BRAF, and CRAF, are proteins that can propagate RAS signaling through the MAPK pathway. RAF activation typically requires membranelocalization, protein conformational changes, and homo- or heterodimerization Without being bound by theory, RAF signaling is a step in melanomagenesis and relates to the ability of pan- RAF inhibitors to limit the growth of each of NRAS- or BRAF-mutant melanoma cells. Disclosed herein is a short-term intervention with a pan-RAF inhibitor to shrink, inhibit, or eliminate early-stage NRAS- and / or BRAF-mutant melanoma precursor lesions. Successful pharmacologic elimination of these precursors can prevent melanoma and reduce the need for some skin biopsies. A similar methodology can prevent the recurrence of surgically excised tumors or reduce the size of congenital nevi.

[0077] Cancer-associated RAF mutants, such as BRAF V600E and K, are constitutively active and function as monomers. These cancer-associated RAF monomers are effectively inhibited by Type I RAF inhibitors, like vemurafenib, dabrafenib, and encorafenib. However, Type I RAF inhibitors can also paradoxically activate the MARK pathway through allosteric activation of the uninhibited RAF molecule in a dimer30. Paradoxical activation of the MAPK pathway is associated with the emergence of secondary RAS-mutant cancers (e.g., squamous-cell carcinomas / SCCs) in patients receiving type I BRAF inhibitors1. Consequently, Type I BRAF inhibitors are now usually combined with downstream MAPK pathway inhibitors to reduce resistance and SCC development.

[0078] A pipeline of preclinical Type II RAF inhibitors has emerged to overcome paradoxical activation of the MAPK pathway (FIG. 2C). Unlike Type I RAF inhibitors, these Type II RAF inhibitors bind and inhibit both protomers in a RAF dimer.

[0079] The standard of care for large congenital nevi (LCN), atypical moles, and early-stage melanomas includes a combination of clinical monitoring (e.g., digital imaging, dermoscopy) and surgical excision (e.g., scalpel, laser, dermabrasion, cryotherapy). However, these approaches have significant drawbacks, including high rates of non-malignant biopsies, locoregional cancer recurrence, and the aesthetic consequences of surgical excision, in addition to surgical site infection, excessive bleeding, delayed w7ound healing, and decreased quality of life during w?ound healing,

[0080] The methods disclosed herein comprise a non-invasive and effective approach to prevent melanoma development and recurrence. For subjects with large congenital nevi, which most often form melanomas within the first ten years of life, the methods disclosed herein limitthe need for surgery and thus significantly reduce surgical risks and consequences, as well as aesthetic concerns.[00811 The targeted cancer therapy disclosed herein primarily blocks signaling through the Mitogen- Activated Protein Kinase (MAPK) pathway. MAPK signaling often begins with RAS (NRAS, KRAS or HRAS) activation of the RAF kinases: ARAF, BRAF, and CRAF Emerging pan-RAF inhibitors, such as exarafenib, block the activities of both mutant and wild-type RAFs, effectively shutting down MAPK signaling13 14. While RAF inhibitors are effective in NRAS- and BRAF-mutant cell lines and pre-clinical models, they have been less successful in early clinical trials due to dose-limiting toxicities and acquired resistance.

[0082] The transition of moles to early-stage melanomas is accompanied by increased MAPK signaling, suggesting a higher dependency on this pathway7Moles and early-stage melanomas have lower genetic and phenotypic heterogeneity, reducing the likelihood of resistance development '. Greater than 80% of moles and melanomas depend on either an NRAS or BRAF mutation. Pan-RAF inhibitors target both NRAS- and BRAF-mutant cancers14, 15. Formulating the pan-RAF inhibitors disclosed herein into a topical formulation, such as for example, a suspension, lotion, ointment, patch, salve, cream, gel, paste, foam, liquid, etc., enables effective application of the therapeutic agent to localized targets, including but not limited to LCN, atypical nevi, and areas of excised early-stage melanomas to prevent cancer development and local recurrence, without systemic effects.

[0083] As an initi l proof of principle described herein, mouse models were developed that replicate the genetic and environmental etiology of human melanoma16 17. These mice express melanocyte-specific oncogenic NRAS or BRAF and develop nevi, which spontaneously form melanoma at a rate comparable to humans. Before the onset of melanoma, NRAS-mutant mice were treated orally with the pan-RAF inhibitor exarafenib for three days. Exarafenib decreased the number of mutant melanocytes in the skin and delayed tumor onset. With topical delivery of the pan-RAF inhibitors disclosed herein, the drug levels are increased in the tumor bearing tissue, relative to drug levels in non-tumor bearing tissue; thus, systemic toxicity is reduced, and overall efficacy is increased.

[0084] As discussed herein, the ability of mutant NRAS to establish melanoma in a mouse is dependent on how well the NRAS mutant engages and activates RAF> MAPK signaling. As shown herein, short-term, preventative oral dosing of pan-RAF inhibitors, e.g. exarafenib ornaporafenib, reduces melanoma risk in an NRAS-mutant mouse melanoma model. Short-term, preventative dosing of an oral pan-RAF inhibitor, e.g. exarafenib, decreases melanin in the skin of NRAS mutant mouse models, suggestive of melanocyte elimination. Topical delivery of pan- RAF inhibitors can limit MAPK signaling and decrease NRAS-mutant melanocyte frequency. Topical pan-RAF inhibitor, e.g. exarafenib, can limit MAPK signaling in the skin of melanoma- prone mice. As shown herein, topical exarafenib, but not dabrafenib, decreases melanin levels in the ear and tail of treated mice. Thus, not all RAF inhibitors are effective preventatives. Topical delivery of dabrafenib, a type I½ RAF inhibitor hastens melanoma development in mice; topical administration of exarafenib, a pan-RAF inhibitor, limits the growth of established NRAS mutant melanomas.II. Compounds

[0085] Disclosed herein are compounds suitable for use in inhibiting, reducing, and / or preventing the initiation or recurrence of a disease or disorder, such as melanoma. In one embodiment, the compound is an inhibitor of the MAPK pathway. In one embodiment, the compound is a pan-RAF inhibitor.

[0086] In one embodiment, the compound is a compound of Formula (I)-(IV) as disclosed herein, including any compound having the structural Formula (I) -(IV) as defined by the variable definitions provided, possible solvates, including hydrates thereof, and amorphous and crystal forms, including one or more polymorphic forms and mixtures thereof. In the case of compounds of Formula (I)-(IV) which possess one or more chiral centers, the compounds may be in the form of a racemic mixture, or one or more isomerically enriched or pure stereoisomers, including enantiomers and diastereomers thereof In such embodiments, compound(s) of Formula (I)-(IV) includes the racemic form as well as the enriched or pure enantiomers and diastereomers. Enantiomerically enriched or pure compounds can be designated using conventional nomenclature, including the designations +, -, R, S, d, I, D and L, according to the predominant isomer present. Where a compound of the invention contains an alkenyl or alkenylene group, cis (E) and trans (Z) isomerism may also occur. In such embodiments, compounds of Formula (I)-(IV) include the individual stereoisomers of the compound of the invention, which can be indicated using conventional, cis / trans nomenclature. It should also be understood that compounds of formula (I) may exist in tautomeric forms otherthan that shown in the formula and alternative tautomeric forms are also included within compounds of Formula (I)-(IV),[00871 A compound of Formula (I)-(IV) (as defined hereinbelow) includes any version, i.e., as the free base or as a pharmaceutically acceptable salt thereof. The compound as any version may be in any form, including amorphous or crystalline forms, specific polymorphic forms, solvates, including hydrates (e.g., mono-, di- and hemi-hydrates), and mixtures of various forms. Intermediates may also be present as salts. Thus, in reference to intermediates, compounds of Formula (I)-(IV) means a compound having that structural formula or a pharmaceutically acceptable salt thereof.

[0088] Representative, non-limiting pharmaceutically active salts include nontoxic acid addition salts and salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.

[0089] In one aspect, a compound is provided comprising a pan-RAF inhibitor.

[0090] In one embodiment, the pan-RAF inhibitor is a compound of Formula (I) or a pharmaceutically acceptable salt, or therapeutically effective derivative or analog thereof:Formul a (I)wherein ring A is a 5- or 6-membered optionally substituted heteroaryl, or 5- or 6- membered optionally substituted heterocyclyl; W is NH, NR7, or O; X is N, C-H, C-F, or C-CN; Y is N, CF, or CH; R is H, CI-C8 optionally substituted alkyl, (C1-C8 optionally substituted alkylene)-OPO(OH)2, C3-C6 optionally substituted cycloalkyl, (C3-C6 optionally substituted cycloalkylene)- OPO(OH)2, C4-C8 optionally substituted cycloalkylalkyl, (C3-C6 optionallysubstituted cycloalkylalkylene)-OPO(OH)2, C3-C6 optionally substituted heterocyclyl, (C3- C6 optionally substituted heterocyclyl )-OPO(OH)2, C3-C6 optionally substituted heterocyclylalkyl, (C3-C6 optionally substituted heterocyclylalkyl)-OPO(OH)2, orCl-C8 optionally substituted alkyl-CO-; R2is H, D or F; R4is halogen, optionally substituted C1-C3 alkyl, -CD3, or optionally substituted C1-C3 alkoxy, R6is H, D, Cl or F; R' is C1-C8 optionally substituted alkyl; or R is not H, and R and R7optionally join to form an optionally substituted heterocyclyl ring;Zis (a)wherein m is 0, 1, 2, or 3; p is 0, 1, 2, 3, or 4; and each R11is independently selected from amino, alkylamino, dialkylamino, -OH, halogen, optionally substituted C1-C6 alkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted C1-C6 alkoxy, optionally substituted C2-C6 alkynyl, optionally substituted -S- alkyl, optionally substituted -SO₂alkyl, optionally substituted C3-C6 cycloalkylalkyl, optionally substituted heterocyclyl, or optionally substituted heterocyclylalkyl; or two R11groups together form an oxo; orwherein m is 0, 1, 2, or 3, n is 0, 1, 2, or 3; p is 0, 1, 2, 3, or 4, and each R13is independently selected from amino, alkylamino. dialkylamino, -OH, halogen, optionally substituted C1-C6 alkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted C1-C6 alkoxy, optionally substituted C2-C6 alkynyl, optionally substituted -S-alkyl, optionally substituted -SO₂alkyl, optionally substituted C3-C6 cycloalkylalkyl, optionally substituted heterocyclyl, or optionally substituted heterocyclylalkyl; or two R11groups together form an oxo Compounds of Formula I can also be found in WO 2021 / 081375 Al.[00911 In one embodiment, the compound is a compound of Formula (II) or a pharmaceutically acceptable salt, or a therapeutically effective derivative or analog thereof:J \ F! HHO. s^.. '• ^,. Xf-. N.H. S A x>- J 6Formula (II).The compound of Formula II is exarafenib.

[0092] In one embodiment, the compound is a compound of Formula (III) or a pharmaceutically acceptable salt, or therapeutically effective derivative or analog thereofFormula (III)or a pharmaceutically acceptable salt thereof, wherein: Z1is O, S, S(=O) or SO2; Z2is N, S or CRa, where Rais H, halo, Cl-4 alkyl or Cl-4 haloalkyl; R1is CN, halo, OH, Cl-4 alkoxy, or C1-4 alkyl that is optionally substituted with one to three groups selected from halo, Cl-4 alkoxy, CN, and hydroxyl, Ring B is selected from phenyl, pyridine, pyrimidine, pyrazine, pyridazine, pyridone, pyrimidone, pyrazinone, pyridazinone, and thiazole, each of which is optionally substituted with up to two groups selected from halo, OH, CN, Cl-4 alkyl, C2-4 alkenyl, -O-(C I -4 alkyl), -NH2, NH-(C1-4 alkyl), -N(C 1 -4 alkyl)2, -SO2R2, NHSO2R2, NHC(O)R2, NHCO2R2, C3-6 cycloalkyl, 5-6 membered heteroaryl -O-C3-6 cycloalkyl, -O-(5-6-membered heteroaryl), C4-8 heterocycloalkyl, and -0-(4-8 membered heterocycloalkyl), where each heterocycloalkyl and heteroaryl contains up to three heteroatoms selected from N, O and S as ring members, where each Cl-4 alkyl, C2-4 alkenyl, C3-8 cycloalkyl 5-6 membered heteroaryl, and 4-8 membered heterocycloalkyl is each optionally substituted with up to three groups selected from oxo, hydroxyl, halo, Cl-4 alkyl, Cl-4 haloalkyl, Cl-4 alkoxy, and -(CH₂)1-2Q where Q is OH, Cl-4 alkoxy, -CN, NH2, -NHR3, -N(R3)2, - SO2R3, NHSO2R3, NHC(O)OR3, or NHC(O)R3; each R2and R3is independently C1-4 alkyl; and Ring B is optionally fused to a 5-6 membered aromatic or nonaromatic ring containing up to two heteroatoms selected from N, O and S, where the 5-6 membered ring can be substituted with halo, C 1-4 alkyl, Cl-4 haloalkyl, or Cl-4 alkoxy,and if the fused ring is non-aromatic the substituent options can further include oxo; each Y is independently selected from Cl -4 alkyl, Cl -4 alkoxy, CN, halo, oxo, -(CH2)pOR4, -(CH2)pN(R4)2, -(CH2)pNHC(O)R4, -(CH2)pNHCOO(Cl-4 alkyl), and imidazole, or two Y groups on Ring A are optionally taken together to form a ring fused to or bridging Ring A, where said fused or bridging ring optionally contains a heteroatom selected from N, O and S as a ring member, and is optionally substituted with up to two groups selected from Cl-4 alkyl, Cl -4 alkoxy, CN, halo, oxo, -(CH2)POR4, -(CH2)PN(R4)2, - (CH2)PNHC(O)R4, and -(CH2)pNHCOO(C1-4 alkyl); each R4is independently H or Cl-4 alkyl; each p is independently 0, 1, or 2, q is 0, 1 or 2; Z3, Z4, and are independently selected from CH and N and optionally NO; L is -C(=O)-NR4-[CY] or -NR4-C(=O)-[CY], where [CY] indicates which atom of L is attached to CY; and CY is an aromatic ring selected from phenyl, pyridine, pyrimidine, pyrazine, pyridazine, pyridone, thiazole, isothiazole, oxazole, pyrazole, and isoxazole, wherein the ring is optionally fused to a thiophene, imidazole, oxazolone, or pyrrole ring; and CY is substituted with up to two groups selected from halo, CN, R3, OR3, SO2R3, -S(=NH)(=O)R3, OH, NH2, NHR5, and -N(R3)2, wherein each R5is independently Cl-4 alkyl, C2-4 alkenyl, C4-6 heterocyclyl, 5- membered heteroaryl containing up to three heteroatoms selected from N, O and S as ring members, or C3-8 cycloalkyl, and R3is optionally substituted with up to four groups selected from oxo, halo, CN, R6, OH, OR6, SO2R6, NH2, NHR6, N(R6)2, NHSO2R6, NHCOOR6, NHC(=0)R6, -CH2OR7, -CH2N(R7)2, wherein each R6is independently Cl-4 alkyl, and each R7is independently H or Cl-4 alkyl; and two R4, R3, R6, or R7on the same nitrogen atom can be taken together to form a 5-6 membered heterocyclic ring optionally containing an additional N, O or S as a ring member and optionally substituted with up to two groups selected from Cl-4 alkyl, oxo, halo, OH, and Cl-4 alkoxy. Compounds of Formula III can also be found inWO 2014 / 151616A1.

[0093] In one embodiment, the compound is a compound of Formula (IV) or a pharmaceutically acceptable salt or a therapeutically effective derivative or analog thereof:Formula (IV).The compound of Formula IV is naporafenib.

[0094] In one embodiment, the pan-RAF inhibitor is AZ628, avutometinib (RO-5126766; CH-5126766; CKI-27; R-7304; RG-7304), belvarafenib (HM95573 / GDC-5573), brimarafenib (BGB-3245), CCT196969, CCT241161, exarafenib (KIN-2787), GNE-9815, IHMT-RAF-128, JZP815 (JSP81538, LY3009120, RAF-265), lifirafenib (BGB-283), LSN3074753, naporafenib, pan-RAF kinase inhibitor 1, RAF709 (QLH11906), SJ-C1044, TBAP-001, tovorafenib (TAK- 520, TAK-580), TAK-632 or a therapeutically effective derivative or analog thereof.

[0095] In one embodiment, the compound is a pan-RAF inhibitor, including, for example Type II RAF inhibitors Examples of pan-RAF inhibitors, such as Type II RAF inhibitors include, but are not limited to, AZ628, belvarafenib, CCT196969, CCT241161, exarafenib, JZP81538, LY3009120, RAF-26536, tovorafenib or therapeutically effective derivatives or analogs thereof (see Table 1 ).Table 1. Exemplary pan-RAF inhibitorsu H H: iCsA. A. X / u 1 J T J 0 "xr-i- v A A.. A. A A O I J. J H - AZ628belvarafenibCt.... F A f " YY-A. / i H r? n H H f' A Av A, A A. N.. M hiN „ Y Y Y rX-'N 0 -AKo Mo f \0 xV. Xv / vji „FHO YF ■'X"HBelvarafenib TFACCT 196969T T / ?O.VNHT0"', NXA.X 1 70 N NCCT241161H £ >z. 1 H 1 *.. Itvy• •! X f | J W' f y; 'N w x i "i >-. S»M" W <-*f- -Z ••• / ::: Z H""TAK 632 1... / F”XF F ^\^yF / RAF-26536R,zv z ( O ) co ■.i 9 v y ''Y H O ZM „ T' YYVNZ VN' A' A ' A XH H ^rtt [xy XH HA A A. I ' Kt " '’X '"' N ''' x''■' N N H M LY3009120?'I I LSN30747531z.t X'~n F / / X.... Z-F O Ov zs Fi""'\,„. / 'p V > X 1 H J) / "■• T HN...ZX....0*. -.z A,.SX.X>N...,,. O r o H r ) AT.■><>U -,. A M *.. A.. z'A VHa.F o X A k,o Lifirafenib (BGB-283) N ' ■''■ R. AF709„ F 0 A / " X....... H k X, A?'kPA ' A-y' 'M' {x'Xx.. J X- H / 'X A VM A ^...„ 1 H H t|?■• X A zk A A\ A zF V..-Q r if o x o ' TA A. A0 FIHMT-RAF-128SJ-C1044

[0096] In another embodiment, the pan-RAF inhibitor is selected from the group of AZ628, belvarafenib, CCT196969, CCT241161, exarafenib, JZP815, and tovorafenib. In yet another embodiment, the pan-RAF inhibitor is exarafenib or naporafenib. In yet another embodiment, the pan-RAF inhibitor is exarafenib.

[0097] In one embodiment, the pan-RAF inhibitor is a paradox breaker, i.e., a compound that interrupts BRAF dimerization through disruption of the aC-helix The compound may be, for example, PLX7904 or PLX8394 or a therapeutically effective derivative or analog thereof:PLX7904 PLX8394

[0098] In yet another embodiment, the pan-RAF inhibitor is a compound that binds to both RAF and MEK proteins, such as for example, a RAF / MEK clamp or molecular glue. In one embodiment, the pan-RAF inhibitor is avutometinib (VS-6766). In another embodiment, a pan-RAF inhibitor NST-628.

[0099] In one embodiment, the compound is exarafenib, the compound of Formula (II). Exarafenib is a highly selective pan-RAF inhibitor that equally inhibits both members of a RAF dimer. In one embodiment, the compound is a therapeutically active derivative or analog of exarafenib.

[0100] In one embodiment, the compound is naporafenib, the compound of Formula (IV). In one embodiment, the compound is a therapeutically active derivative or analog of naporafenib.

[0101] In one embodiment, the compound is avutometinib. In one embodiment, the compound is a therapeutically active derivative or analog of avutometinib.

[0102] In one embodiment, the compound has an IC50 of less than about 100 nM, less than about 50 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, or less than about 1 nM.

[0103] In one embodiment, the compound may be a prodrug of any compound disclosed herein, for example, a prodrug of a pan-RAF inhibitor, such as a compound of Formula (I)-(IV).III. Pharmaceutical Compositions

[0104] A pharmaceutical composition described herein may be made in any manner not inconsistent with the technical objectives of the current disclosure.

[0105] In one aspect, a pharmaceutical composition is provided comprising any compound disclosed herein and at least one pharmaceutically acceptable carrier or excipient. In embodiment, the composition comprises a pan-RAF inhibitor from Section II.

[0106] In one embodiment, the pharmaceutical composition is provided as a dermal formulation. The formulation for dermal administration may be, for example,, a suspension, lotion, ointment, spray, patch, salve, cream, gel, paste, foam, liquid, or the like.

[0107] In one embodiment, the pharmaceutical composition further comprises at least one therapeutic agent, for example, PF- 07284892 / ARRY-558 or a therapeutically active analog thereof.

[0108] Also provided herein are pharmaceutical compositions comprising the compounds disclosed herein in Section II and one or more pharmaceutically acceptable carriers, excipients, buffers, adjuvants, stabilizers, or other materials.

[0109] Suitable pharmaceutical carriers and their formulations are described in Remington’s Pharmaceutical Sciences (Mack Publishing Co., Easton, 19th ed. 1995). Preferredpharmaceutical carriers depend upon the intended mode of administration of the active agent. The pharmaceutical compositions can be in any form suitable for oral, parenteral, topical, intranasal, ophthalmic, otic, rectal, intra-vaginal, or transdermal administration.

[0110] Representative, non-limiting examples of excipients include binders, disintegrates, superdisintegrants, lubricants, diluents, fillers, flavors, glidants, sorbents, solubilizers, chelating agents, emulsifiers, thickening agents, dispersants, stabilizers, suspending agents, adsorbents, granulating agents, preservatives, buffers, coloring agents and sweeteners or combinations thereof. In one embodiment, the pharmaceutical composition is formulated for non-systemic administration. In one embodiment, the composition is formulated for dermal administration, which may include topical application to the dermis. In another embodiment, the composition is formulated for administration intradermally, interdennally, transdermally, or the like,

[0111] In one embodiment, the pharmaceutical composition is a topical formulation, e g., a topical solution, a suspension, a cream, an ointment, a lotion, a foam, a patch or a gel.

[0112] The concentration of the pan-RAF inhibitor in the topical composition may vary. In one embodiment, the pan-RAF inhibitor is present at a concentration of less than about 0.1%, less than about 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03% or less than 0.02% based on the total weight of the formulation. In one embodiment, the pan-RAF inhibitor is present at a concentration from about 0.005% to 0.05%, 0.008% to 0,04%, 0.01% to 0.03% or about 0.025% based on the total weight of the formulation. In another embodiment, the pan-RAF inhibitor is present at a concentration from about 0.1% w / w to about 5% w / w or about 1% w / w to about 4% w / w, based on the total weight of the formulation.

[0113] The dose administered may be any suitable non-systemic dose. In one embodiment, the pharmaceutical composition or combination disclosed herein can be in unit dosage of about 1-1000 mg of active ingredient(s) for a subject of about 50-70 kg, or about 1-500 mg or about 1-250 mg or about 1- 150 mg or about 0.5-100 mg, or about 1-50 mg of active ingredients. The therapeutically effective dosage of a compound, the pharmaceutical composition, or the combinations thereof, is dependent on the species of the subject, the body weight, age and individual condition, the disorder or disease or the severity thereof being treated.IV. Methods of Use

[0114] In another aspect, disclosed herein are methods of using the compounds and pharmaceutical compositions disclosed herein.

[0115] In one aspect, disclosed herein is a method of reducing the size of a melanoma precursor lesion in a subject in need thereof, the method comprising: non-systemically administering a pan-RAF inhibitor to the melanoma precursor lesion. In one embodiment the pan-RAF inhibitor inhibits a MAP kinase pathway. In one embodiment, the method does not comprise co-administering a downstream ATP competitive inhibitor of the MAP kinase pathway. In a still further embodiment, the melanoma precursor lesion has not been diagnosed as melanoma. In one embodiment, the melanoma precursor lesion has not been pathologically diagnosed as melanoma

[0116] In one embodiment, the size of the melanoma precursor lesion is reduced by 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20% or 10%. In another embodiment, the size of the melanoma precursor lesion is reduced by 90%-10%, 80%-10%, 70%-10%, 60%-10%, 50%- 10%, 40%-10%, 30%-10%, 20%-10%, 90%-20%, 80%-20%, 70%-20%, 60%-20%, 50%-20%, 40%- 20%, 30%-20%, 90%-30%, 80%-30%, 70%-30%, 60%-30%, 50%-30%, 40%-30%, 90%-40%, 80%-40%, 70%-40%, 60%-40%, or 50%-40%. In another embodiment, reducing the size of the melanoma precursor lesion comprises eliminating the melanoma precursor lesion.

[0117] In another aspect, disclosed herein is a method of inhibiting progression of a melanoma precursor lesion to melanoma in a subject in need thereof, the method comprising: non-systemically administering a pan-RAF inhibitor to the melanoma precursor lesion. In one embodiment the pan-RAF inhibitor inhibits a MAP kinase pathway. In one embodiment, the method does not comprise co-administering a downstream ATP competitive inhibitor of the MAP kinase pathway. In a still further embodiment, the melanoma precursor lesion has not been diagnosed as melanoma. In one embodiment, the melanoma precursor lesion has not been pathologically diagnosed as melanoma.

[0118] In one embodiment, inhibiting progression comprises not forming a benign nevus, or not forming a dysplastic nevus, or inhibiting a dysplastic nevus from developing into melanoma. In another embodiment, inhibiting progression inhibits Stage 0 melanoma.

[0119] In another aspect, disclosed herein is a method of preventing melanoma in a subject in need thereof, the method comprising: non-systemically administering a pan-RAF inhibitor tothe subject In one embodiment, the pan-RAF inhibitor is administered to one or more melanoma precursor lesions. In one embodiment the method does not comprise co-administering a downstream ATP competitive inhibitor of the MAP kinase pathway. In a further embodiment, the subject does not have melanoma. In one embodiment, the melanoma precursor lesion is not melanoma. In another embodiment, there is no pathological melanoma diagnosis for the melanoma precursor lesion. In yet another embodiment, the subject does not have a current melanoma diagnosis.

[0120] In one embodiment of any method disclosed herein, the subject: (a) was born with large congenital nevi; or (b) has atypical or multiple nevus syndrome. In another embodiment, the subject does not have melanoma. In another embodiment, the subject has not had a melanoma diagnosis for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 years. In yet another embodiment, the subject has never been diagnosed with melanoma. In yet another embodiment of any method disclosed herein, the subject: (a) was born with large congenital nevi; and / or (b) has atypical or multiple nevus syndrome.

[0121] In one embodiment, the pan-RAF inhibitor is administered for no more than 12 weeks, 11 weeks, 10 weeks, 9 weeks, 8, weeks, 7 weeks, 6 weeks, no more than 5 weeks, no more than 4 weeks, no more than 3 weeks, no more than 2 weeks, or no more than 7 days. In another embodiment, the pan-RAF inhibitor is administered for 12 weeks, 11 weeks, 10 weeks, 9 weeks, 8, weeks, 7 weeks, 6 weeks, 5 weeks, 4 weeks, 3 weeks, 2 weeks or 7 days. In another embodiment, the pan-RAF inhibitor is administered until the prophylactic goal(s) are achieved. In yet another embodiment, the pan-RAF inhibitor is administered until one or more negative side effects outweigh the therapeutic benefit of the pan-RAF inhibitor. In yet another embodiment, the pan-RAF inhibitor is administered until therapeutic resistance is observed.

[0122] In one embodiment of any method disclosed herein, the melanoma precursor lesion has not been biopsied. In another embodiment, the melanoma precursor lesion has not been incised or excised. In yet another embodiment, the melanoma precursor lesion comprises a cluster of melanocytes. In a further embodiment, the melanoma precursor lesion is a dysplastic nevus.

[0123] In another embodiment of any method disclosed herein, the melanoma precursor lesion comprises cells having an NRAS and / or BRAF mutation.

[0124] The NRAS mutation may be any NRAS mutation, including, for example a mutation affecting codons 12, 13, or 61, Without being bound by any particular theory, it is believed that melanoma formation is dependent upon substitution- and isoform-specific structural differences in NRAS that enhance RAF dimerization and MAPK> ERK signaling, i.e., NRAS mutants differentially recognize and promote RAF activation.

[0125] In one embodiment, the NRAS mutation is in codon 61. In another embodiment, the NRAS mutation is Q61R (Glutamine to Arginine), Q61K (Glutamine to Lysine), Q61L (Glutamine to Leucine), Q61H (Glutamine to Histidine), or Q61P (Glutamine to Proline).

[0126] In one embodiment, the methods disclosed herein have a differential impact (e.g., greater reduction in the number or size of nevi or greater impact on melanoma development) on an NRAS codon 61 mutation-specific basis. That is, the method shows improved results based on a particular mutation selected from those listed above or known in the art

[0127] In one embodiment, the NRAS mutation is in codon 12. In another embodiment, the NRAS mutation is G12D (Glycine to Aspartic acid), G12V (Glycine to Valine); G12C (Glycine to Cysteine); G12S (Glycine to Serine); G12A (Glycine to Alanine); G12R (Glycine to Arginine).

[0128] In one embodiment, the methods disclosed herein have a differential impact (e.g., greater reduction in number or size of nevi or greater impact on melanoma development) on an NRAS codon 12 mutation-specific basis. That is, the method shows improved results based on a particular mutation selected from those listed above or known in the art.

[0129] In one embodiment, the NRAS mutation is in codon 13. In another embodiment, the NRAS mutation is G13D (Glycine to Aspartic acid): G13R (Glycine to Arginine).

[0130] In one embodiment, the methods disclosed herein have a differential impact (e.g., greater reduction in number or size of nevi or greater impact on melanoma development) on an NRAS codon 13 mutation-specific basis. That, is, the method shows improved results based on a particular mutation selected from those listed above or known in the art.

[0131] In one embodiment, the BRAF mutation is a point mutation at residues 594, 600, or 601. In another embodiment, the BRAF point mutation is a V600E (V aline to Glutamic acid), V600K (Valine to Lysine), V600R (Valine to Argine), V600D (Valine to Aspartic acid), V600M (Valine to Methionine), V600G (Valine to Glycine), K601E (Lysine to Glutamic acid), or D594N (Aspartic acid to Asparagine) substitution.

[0132] In one embodiment, the melanoma precursor lesion comprises cells having an NRAS mutation at codon 12, 13, or 61, or a BRAF point mutation at residue 594, 600 or 601. In another embodiment, the NRAS mutation is Q61R, Q61K, Q61L, Q61H, or Q61P. In yet another embodiment, the NRAS mutation is G12D, G12V, G12C, G12S, G12Aor G12R. In still a further embodiment, the NRAS mutation is GI3D or G13R. In yet another embodiment, the BRAF mutation is V600E, V600K, V600R, V600D, V600M, V600G, K601E, or D594N.

[0133] In another aspect, disclosed herein is a method for inhibiting recurrence of melanoma cells in a border region remaining after excision of a local melanoma in a subject, the method comprising: non-systemically administering a pan-RAF inhibitor to the border region of the subject following the excision of the local melanoma; wherein the method does not comprise co¬ administering a downstream ATP competitive inhibitor of the MAP kinase pathway. In one embodiment, the local melanoma is a Stage 0, Stage I or Stage II melanoma. In another embodiment, the subject has not been re-diagnosed with melanoma at the site of the local melanoma excision following the excision and prior to non-systemically administering the pan-RAF inhibitor to the border region.

[0134] In another embodiment of any method disclosed herein, the subject has not had a melanoma diagnosis for at least one, two, three, four or five years.

[0135] In another embodiment of any method disclosed herein, the pan-RAF inhibitor inhibits two or more of A-RAF, B-RAF and C-RAF. In yet another embodiment, the pan-RAF inhibitor inhibits both protomers of any RAF dimer. In yet another embodiment, the pan-RAF inhibitor is a RAF / MEK inhibitor. In yet another embodiment, the pan-RAF inhibitor is a paradox breaker.

[0136] In another embodiment of any method disclosed herein, the subject has not been previously treated with a pan-RAF inhibitor, topically or systemically. In another embodiment, the subject has been previously treated topically with a pan-RAF inhibitor, but the site / lesion / nevus being treated has not been previously treated topically with a pan-RAF inhibitor. In another embodiment, the subject / site / lesion / nevus has been previously treated topically with a pan-RAF inhibitor.

[0137] In one embodiment of any method disclosed herein, the pan RAF inhibitor is AZ628, avutometinib, belvarafenib, brimarafenib, CCT 196969, CCT241161, exarafenib, GNE-9815, IHMT-RAF-128, JZP815, lifirafenib, LSN3074753, naporafenib, pan-RAF kinase inhibitor 1,RAF709, SJ-C1044, TBAP-001, tovorafenib, TAK-632 or a therapeutically effective derivative or analog thereof. In another embodiment of any method disclosed herein, the pan-RAF inhibitor is selected from the group of AZ628, belvarafenib, CCT196969, CCT241161, exarafenib, JZP815, and tovorafenib. In another embodiment of any method disclosed herein, the pan-RAF inhibitor is exarafenib or naporafenib. In another embodiment of any method disclosed herein, the pan-RAF inhibitor is exarafenib. In yet another embodiment of any method disclosed herein, the pan-RAF inhibitor is avutometinib.

[0138] In one embodiment, the pan-RAF inhibitor is co-administered with an inhibitor of RAS, an inhibitor of RTK, and / or an inhibitor of Shp2.

[0139] In one embodiment of any method disclosed herein, the method does not comprise systemic administration of a therapeutic agent for the treatment of cancer; in one embodiment, the cancer is melanoma

[0140] In one embodiment of any method disclosed herein, the method does not comprise co-administering an ATP competitive MEK inhibitor. In yet another embodiment, the method does not comprise co-administering a MEK inhibitor. In another embodiment, the method does not comprise co-administering an ATP competitive MEK inhibitor or a non-ATP competitive MEK inhibitor. In one embodiment, the method does not comprise co-administering an inhibitor downstream of RAF. In another embodiment, the method does not comprise co-administering an inhibitor of any step of the MAP kinase pathway. In yet another embodiment, the method does not comprise co-administering an inhibitor of one or more of RAS, MEK, ERK. In yet another embodiment, the method does not comprise co-administering a mitochondrial inhibitor. In another embodiment, the method does not comprise co-administering a mitochondrial respiration inhibitor. In another embodiment, the method does not comprise co-administering an oxidative phosphorylation inhibitor.

[0141] In one aspect, a method is provided for reducing or eliminating melanoma precursor lesions comprising administering a therapeutically effective amount of a compound or pharmaceutical composition disclosed herein to a subject in need thereof, thereby reducing or eliminating melanoma precursor lesions. In one embodiment, the melanoma precursor lesion comprises an NRAS- and / or BRAF-mutant lesion predisposed to melanoma. In one embodiment, the subject is an animal model of melanoma or a human subject.

[0142] Optionally, the method may further comprise administering at least one additional therapeutic agent (e.g., a MEK inhibitor, an upstream Shp2 inhibitor). In one embodiment, the method does not comprise administering a second therapeutic agent for reducing or eliminating melanoma precursor lesions.

[0143] The percentage of reduction may vary, in one embodiment, the percentage of reduction in melanoma precursor lesions is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% compared to a control. Reduction can be measured by any suitable method, e.g, imaging, dermoscopy.

[0144] In another aspect, a method is provided for reducing a melanoma precursor progression rate comprising administering a therapeutically effective amount of a compound or pharmaceutical composition disclosed herein to a subject in need thereof, reducing a melanoma precursor lesion progression rate. In one embodiment, the melanoma precursor lesion comprises an NRAS- and / or BRAF-mutant lesion predisposed to melanoma. In one embodiment, the subject is an animal model of melanoma or a human subject. Optionally, the method may further comprise administering at least one additional therapeutic agent (e.g., a MEK inhibitor, an upstream Shp2 inhibitor). In one embodiment, the method does not comprise administering a second therapeutic agent for reducing a melanoma precursor progression rate.

[0145] The percentage of progression rate reduction may vary. In one embodiment, the percentage of reduction is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% compared to a control. Reduction can be measured by any suitable method, for example, histopathological analysis or dermoscopy.

[0146] In yet another aspect, a method is provided for reducing the size of melanoma precursor lesions comprising administering a therapeutically effective amount of a compound orpharmaceutical composition disclosed herein to a subject in need thereof, thereby reducing the size of melanoma precursor lesions. In one embodiment, the melanoma precursor lesion comprises an NRAS- and / or BRAF-mutant lesion predisposed to melanoma. In one embodiment, the subject is an animal model of melanoma or a human subject. Optionally, the method may further comprise administering at least one additional therapeutic agent. In one embodiment, the method does not comprise administering second therapeutic agent for reducing the size of melanoma precursor lesions.

[0147] The percentage of size reduction may vary. In one embodiment, the percentage of reduction is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% reduced compared to a control. In one embodiment, the percentage reduction is between about 10% and about 20%, between about 20% and about 30%, between about 30% and about 40%, between about 40% and about 50%, between about 50% and about 60%, between about 60% and about 70%, between about 70% and about 80%, between about 80% and about 90%, or between about 90% and about 100% compared to a control. The percentage reduction may be measured by any suitable method known in the art, such as imaging or measuring.

[0148] In yet another aspect, a method is provided for reducing, inhibiting, and / or preventing melanoma, melanocytic lesions, and / or residual tumor cells after local melanoma excision, the method comprising administering a compound or pharmaceutical composition disclosed herein to a subject in need thereof, thereby reducing, inhibiting, and / or preventing melanoma, melanocytic lesions, and / or residual tumor cell expansion after local melanoma excision. In one embodiment, the melanoma precursor lesion comprises an NRAS- and / or BRAF-mutant lesion predi sposed to melanoma. In one embodiment, the subject is an animal model of melanoma, a 3D skin reconstruct, or a human subject. Optionally, the method may further comprise administrating at least one additional therapeutic agent. In one embodiment, the method does not comprise administering a second therapeutic agent for reducing, inhibiting, and / or preventing melanoma, melanocytic lesions, and / or residual tumor cells after local melanoma excision.

[0149] Reduction can be measured by any suitable method, such as imaging, measuring, histopathological analysis or melanoma-free survival.

[0150] In any of the aspects described herein, the subject may be a human subject at risk for melanoma. Risk assessment may be determined, for example, by clinical presentation of the melanoma precursor lesion, personal history, phenotypic traits, genetic traits, environmental exposure, or any combination thereof.

[0151] In one embodiment, the method is administered to a population of subjects, e.g., a population of human subjects in a clinical trial. According to this embodiment, the results of the method may be measured across the population of subjects.

[0152] In one embodiment, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% of human subjects exhibit a response to treatment with a compound or composition disclosed herein.

[0153] In one embodiment, a method is provided for selectively inducing apoptosis in melanoma cells via non-systemic administration of a pan-RAF inhibitor. In another embodiment, a method is provided for non-systemically administering a pan-RAF inhibitor and inducing senescence in cells, including cells having an NRAS and / or BRAF mutation. In yet another embodiment, a method is provided for inducing immune-mediated clearance of cells, including cells having an NRAS and / or BRAF mutation, via non-systemic administration of a pan-RAF inhibitor. In a variation of any embodiment, the method does not include administration of a second therapeutic agent.

[0154] Although not wishing to be bound by any theory, it is believed that transiently inhibiting RAF kinases may contribute to the results of the methods described herein.

[0155] The subject according to the embodiments disclosed herein may be any suitable subject, e.g., a mammal or human. In one embodiment, the subject is a dog, for example, a dog at risk of developing melanoma.

[0156] In one embodiment, the subject is an animal model of melanoma. In one embodiment, more than one, e.g., a group of 2, 3, 4, 5, 6, 7, 8, 9 or 10 or more animals may be treated.According to this embodiment, reduction or prevention can be measured across a population of subjects.

[0157] In one embodiment, the subject is a human in a clinical trial. According to this embodiment, reduction can be measured across a population of subjects, e.g., a group of humans in a clinical trial.

[0158] In one embodiment, the subject is a human being treated by a physician or other health care provider.

[0159] In one embodiment, the subject is a human at risk for melanoma, for example, a subject that does not currently have melanoma or has not been previously diagnosed with melanoma, such as local melanoma. The risk may be determined, for example, phenotypically (e g, fair skin, age, having many common nevi, large nevi, multiple nevi, atypical nevi), genetically (e.g, family history, melanocortin-1 receptor (MC1R) gene variants) or on the basis of occupational or other exposures (e.g., significant sun exposure). In one embodiment, the subject is considered to be at high risk, having previously had a melanoma lesion that was excised.

[0160] The compound or composition disclosed herein may be administered one or more times to the subject(s). In one embodiment, the compositions are administered 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times or more.

[0161] In one embodiment, the cancer targeted, treated, reduced, inhibited, and / or prevented according to the methods disclosed herein is a cancer other than melanoma. As used herein the term “cancer” refers to a broad group of diseases characterized by the uncontrolled growth of abnormal cells in the body. A "cancer" or "cancer tissue" can include a tumor.

[0162] As disclosed herein, administration of a therapeutic, such as a pan-RAF inhibitor can continue for as long as medically indicated or until a desired therapeutic effect (e.g., those described herein) is achieved For example, patients can be treated until complete response (e.g. all treated melanoma precursor lesions have appropriately reduced in size or disappeared. It is anticipated that non-systemic administration of the pan-RAF inhibitor is continued for at least 12 weeks, 11 weeks, 10 weeks, 9 weeks, 8 weeks, 7 weeks, 6 weeks, 5 weeks, 4 weeks, 3 weeks, 2 weeks, or 1 week from the time of initial dose provided that the subject has no evidence of clinically significant deterioration of health status requiring discontinuation of treatment and isable to tolerate the treatment. However, the course of treatment for any individual patient can be modified in clinical practice.

[0163] In one embodiment, a cancer is selected from any disease where uncontrolled cellular proliferation occurs. A representative but non-limiting list of cancers that the disclosed compositions can be used to treat, inhibit, reduce, and / or prevent include melanoma, colorectal cancers, thyroid cancers, biliary tract cancers, endometrial cancers, lung cancers (including, but not limited to non-small cell lung carcinoma and adenocarcinoma), myeloid leukemias (including, but not limited to acute myeloid leukemia (AML) and / or chronic myeloid leukemias (CML)), brain cancers (including gliomas), bladder cancers, multiple myeloma.Rhabdomyosarcoma, stomach cancer, esophageal cancer, and ovarian cancers.

[0164] In one optional embodiment, a method disclosed herein further comprises administrating at least one additional therapeutic agent to the subject in need thereof The additional therapeutic agent may vary. In one embodiment, the additional optional therapeutic agent is selected from the group consisting of a MEK inhibitor or an upstream Shp2 inhibitor. In another embodiment, neither a MEK inhibitor nor an upstream Shp2 inhibitor is administered as an additional therapeutic agent.

[0165] Representative, non-limiting examples of MEK inhibitors include ARRY-142886 (6-(4-bromo-2-chloro-phenylamino)-7-f!uoro-3-methyl-3H- benzoimidazole-5-carbox lic acid (2-hydroxy-ethoxy)-amide, Array BioPharma), ARRY-438162 (Array BioPharma), AS- 1940477 (Astellas), AS-703988 (Merck KGaA), bentamapimod (Merck KGaA), BI-847325 (Boehringer Ingelheim), E-6201 (Eisai), GDC-0623 (Hoffmann-La Roche), cobimetinib (GDC-0973, Hoffmann-La Roche), L783277 (Merck), MEK162 (Array BioPharma), PD 098059 (2-(2'-amino-3'-methoxphenyl)~ oxanap hthalen-4-one, Pfizer), PD 184352 (CI-1040, Pfizer), PD-0325901 (Pfizer), pimasertib (Santhera Pharmaceuticals), RDEA119 (Ardea Biosciences / Bayer), refametinib (AstraZeneca), RG422 (Chugai Pharmaceutical Co.), R0092210 (Roche), R04987655 (Hoffmann-La Roche), selumetinib (AZD6244) (AstraZeneca), SL327 (Sigma), TAK-733 (Takeda), trametinib (Japan Tobacco), U0126 (l,4-diamino-2,3-dicyano-l,4-bis(2-aminophenylthiojbutadiene, Sigma), WX-554 (Wilex), pharmaceutically acceptable salts thereof, and combinations thereof.

[0166] Other therapeutic agents that can be used in combination with the compounds and pharmaceutical compositions disclosed herein include, but are not limited to, Abemaciclib,Abiraterone Acetate, ABITREXATE® (Methotrexate), ABRAXANE® (Paclitaxel Albumin-stabilized Nanoparticle Formulation), ABVD, ABVE, ABVE-PC, AC, AC-T, ADCETRIS® (Brentuximab Vedotin), ADE, Ado-Trastuzumab Emtansine, ADRIAMYCIN® (Doxorubicin Hydrochloride), Afatinib Dimaleate, AFINITOR® (Everolimus), AKYNZEO® (Netupitant and Palonosetron Hydrochloride), ALDARA® (Imiquimod), Aldesleukin, ALECENSA® (Alectinib), Alectinib, Alemtuzumab, ALIMTA® (Pemetrexed Disodium), ALIQOPA® (Copanlisib Hydrochloride), ALKERAN™ for Injection (Melphalan Hydrochloride), ALKERAN™ Tablets (Melphalan), ALOXI® (Palonosetron Hydrochloride), ALUNBRIG® (Brigatinib), AMBOCHLORIN® (Chlorambucil), AMBOCLORIN® (Chlorambucil), Amifostine, Aminolevulinic Acid, Anastrozole, Aprepitant, AREDIA® (Pamidronate Disodium), ARIMIDEX® (Anastrozole), AR0MAS1N® (Exemestane), ARRANON® (Nel arabine), Arsenic Trioxide, ARZERRA® (Ofatumumab), Asparaginase Erwinia chrysanthemi, Atezolizumab, AVASTIN® (Bevacizumab), Avelumab, Axitinib, Azacitidine, BAVENCIO® (Avelumab), BEACOPP, BECENUM® (Carmustine), BELEODAQ® (Belinostat), Belinostat, Bendamustine Hydrochloride, BEP, BESPONSA® (Inotuzumab Ozogamicin), Bevacizumab, Bexarotene, BEXXAR® (Tositumomab and Iodine I13’ Tositumomab), Bicalutamide, BICNU* (Carmustine), Bleomycin, Blinatumomab, BLINCYTO® (Blinatumomab), Bortezomib, BOSULIF® (Bosutinib), Bosutinib, Brentuximab Vedotin, Brigatinib, BuMel, Busulfan, BUSULFEX® (Busulfan), Cabazitaxel, CABOMETYX® (Cabozantinib-S-Malate), Cabozantinib-S-Malate, CAF, CAMPATH® (Alemtuzum b), CAMPTOSAR® (Irinotecan Hydrochloride), Capecitabine, CAPOX, CARAC® (Fluorouracil-Topical), Carboplatin, CARBOPLATIN-TAXOL, Carfilzomib, CARMUBRIS® (Carmustine), Carmustine, Carmustine Implant, CASODEX® (Bicalutamide), CEM, Ceritinib, CERUBIDINE® (Daunorubicin Hydrochloride), CERVARIX® (Recombinant HPV Bivalent Vaccine), Cetuximab, CEV, Chlorambucil, CHLORAMBUCIL¬ PREDNISONE, CHOP, Cisplatin, Cladribine, CLAFEN® (Cyclophosphamide), Clofarabine, CLOFAREX® (Clofarabine), CLOLAR® (Clofarabine), CMF, Cobimetinib, COMETRIQ® (Cabozantinib-S-Malate), Copanlisib Hydrochloride, COPDAC, COPP, COPP-AB V, COSMEGEN® (Dactinomycin), COTELLIC® (Cobimetinib), Crizotinib, CVP, Cyclophosphamide, CYFOS® (Ifosfamide), CYRAMZA® (Ramucirumab), Cytarabine, Cytarabine Liposome, CYTOSAR-U® (Cytarabine), CYTOXAN® (Cyclophosphamide), Dabrafenib, Dacarbazine, DACOGEN® (Decitabine), Dactinomycin, Daratumumab,DARZALEX® (Daratumumab), Dasatinib, Daunorubicin Hydrochloride, Daunorubicin Hydrochloride and Cytarabine Liposome, Decitabine, Defibrotide Sodium, DEFITELIO® (Defibrotide Sodium), Degarelix, Denileukin Diftitox, Denosumab, DEPOCYT® (Cytarabine Liposome), Dexamethasone, Dexrazoxane Hydrochloride, Dinutuximab, Docetaxel, DOXIL® (Doxorubicin Hydrochloride Liposome), Doxorubicin Hydrochloride, Doxorubicin Hydrochloride Liposome, DOX-SL® (Doxorubicin Hydrochloride Liposome), DTIC-DOME® (Dacarbazine), Durvalumab, EFUDEX® (Fluorouracil— Topical), ELITEK® (Rasburicase), ELLENCE® (Epirubicin Hydrochloride), Elotuzumab, ELOXATIN® (Oxaliplatin), Eltrombopag Olamine, EMEND® (Aprepitant), EMPLICITI® (Elotuzumab), Enasidenib Mesylate, Enzalutamide, Epirubicin Hydrochloride, EPOCH, ERBITUX® (Cetuximab), Eribulin Mesylate, ERIVEDGE® (Vismodegib), Erlotinib Hydrochloride, ERWINAZE® (Asparaginase Erwinia chrysanthemi), ETHYOL® (Amifostine), Etopophos ETOPOPHOS® (Etoposide Phosphate), Etoposide, Etoposide Phosphate, EVACET® (Doxorubicin Hydrochloride Liposome), Everolimus, EVISTA® (Raloxifene Hydrochloride), EVOMELA® (Melphalan Hydrochloride), Exemestane, 5-FU® (Fluorouracil Injection), 5-FU® (Fluorouracil— Topical), FARESTON® (Toremifene), FARYDAK® (Panobinostat), FASLODEX® (Fulvestrant), FEC, FEMARA® (Letrozole), Filgrastim, FLUDARA® (Fludarabine Phosphate), Fludarabine Phosphate, FLUOROPLEX® (Fluorouracil— Topical), Fluorouracil Injection, Fluorouracil— Topical, Flutamide, FOLEX® (Methotrexate), FOLEX PFS® (Methotrexate), FOLFIRI, FOLFIRI-BEVACIZUMAB, FOLFIRI-CETUXIMAB, FOLFERINOX, FOLFOX, FOLOTYN® (Pralatrexate), FU-LV, Fulvestrant, GARDASIL® (Recombinant HPV Quadrivalent Vaccine), GARDASIL 9® (Recombinant HPV Nonaval ent Vaccine), GAZYVA® (Obinutuzumab), Gefitinib, Gemcitabine Hydrochloride, GEMCITABINE-CISPLATIN, GEMCITABINEOXALIPLATIN, Gemtuzumab Ozogamicin, GEMZAR® (Gemcitabine Hydrochloride), GILOTRIF* (Afatinib Dimaleate), GLEEVEC® (Imatinib Mesylate), GLIADEL® (Carmustine Implant), GLIADEL WAFER® (Carmustine Implant), Glucarpidase, Goserelin Acetate, HALAVEN® (Eribulin Mesylate), HEMANGEOL® (Propranolol Hydrochloride), HERCEPTIN® (Trastuzumab), HPV Bivalent Vaccine, Recombinant, HPV Nonavalent Vaccine, Recombinant, HPV Quadrivalent Vaccine, Recombinant, HYCAMTIN® (Topotecan Hydrochloride), HYDREA® (Hydroxyurea), Hydroxyurea, Hyper-CVAD, IB RANCE® (Palbociclib), Ibritumomab Tiuxetan, Ibrutinib, ICE, ICLUSIG® (Ponatinib Hydrochloride),IDAMYCIN® (Idarubicin Hydrochloride), Idarubicin Hydrochloride, Idelalisib, IDHIFA® (Enasi denib Mesylate), IFEX® (Ifosfamide), Ifosfamide, IFOSFAMIDUM® (Ifosfamide), IL-2 (Aldesleukin), Imatinib Mesylate, IMBRUVICA® (Ibrutinib), IMFINZI® (Durvalumab), Imiquimod, IMLYGIC® (Talimogene Laherparepvec), INLYTA® (Axitinib), Inotuzumab Ozogamicin, Interferon Alfa-2b, Recombinant, Interleukin-2 (Aldesleukin), INTRON A® (Recombinant Interferon Alfa-2b), Iodine 1 131 Tositumomab and Tositumomab, Ipilimumab, IRESSA® (Gefitinib), Irinotecan Hydrochloride, Irinotecan Hydrochloride Liposome, ISTODAX® (Romidepsin), Ixabepilone, Ixazomib Citrate, IXEMPRA® (Ixabepilone), JAKAFI® (Ruxolitinib Phosphate), JEB, JEVTANA® (Cabazitaxel), KADCYLA® (Ado-Trastuzumab Emtansine), KEOXIFENE® (Raloxifene Hydrochloride), KEPIVANCE® (Palifermin), KEYTRUDA® (Pembrolizumab), KISQALI® (Ribociclib), KYMRIAH® (Tisagenlecleucel), KYPROLIS® (Carfilzomib), Lanreotide Acetate, Lapatinib Ditosylate, LARTRUVO® (Olaratumab), Lenalidomide, Lenvatinib Mesylate, LENVTMA®’ (Lenvatinib Mesylate), Letrozole, Leucovorin Calcium, LEUKERAN® (Chlorambucil), Leuprolide Acetate, LEUSTATIN® (Cladribine), LEVULAN® (Aminolevulinic Acid), LINFOLIZIN® (Chlorambucil), LIPODOX® (Doxorubicin Hydrochloride Liposome), Lomustine, LONSURF® (Trifluridine and Tipiracil Hydrochloride), LUPRON® (Leuprolide Acetate), LUPRON DEPOT® (Leuprolide Acetate), LUPRON DEPOT-PED® (Leuprolide Acetate), LYNPARZA® (Olaparib), MARQIBO® (Vincristine Sulfate Liposome), MATULANE® (Procarbazine Hydrochloride), Mechlorethamine Hydrochloride, Megestrol Acetate, Melphalan, Melphalan Hydrochloride, Mercaptopurine, Mesna, MESNEX® (Mesna), METHAZOLASTONE® (Temozolomide), Methotrexate, METHOTREXATE LPF® (Methotrexate), Methylnaltrexone Bromide, MEXATE® (Methotrexate), MEXATE-AQ® (Methotrexate), Midostaurin, Mitomycin C, Mitoxantrone Hydrochloride, MITOZYTREX® (Mitomycin C), MOPP, MOZOBIL® (Plerixafor), MUSTARGEN® (Mechlorethamine Hydrochloride), MUTAMYCIN® (Mitomycin C), MYLERAN® (Busulfan), MYLOSAR® (Azacitidine), MYLOTARG® (Gemtuzumab Ozogamicin), NANOPARTICLE PACLITAXEL® (Paclitaxel Albumin-stabilized Nanoparticle Formulation), NAVELBINE® (Vinorelbine Tartrate), Necitumumab, Nelarabine, NEOSAR® (Cyclophosphamide), Neratinib Maleate, NERLYNX® (Neratinib Maleate), Netupitant and Palonosetron Hydrochloride, NEULASTA® (Pegfilgrastim), NEUPOGEN® (Filgrastim), NEXAVAR® (Sorafenib Tosylate), NILANDRON® (Nilutamide), Nilotinib, Nilutamide,NINLARO® (Ixazomib Citrate), Niraparib Tosylate Monohydrate, Nivolumab, NOLVADEX® (Tamoxifen Citrate), NPLATE® (Romiplostim), Obinutuzumab, ODOMZO® (Sonidegib), OEPA, Ofatumumab, OFF, Olaparib, Olaratumab, Omacetaxine Mepesuccinate, ONCASPAR® (Pegaspargase), Ondansetron Hydrochloride, ONIVYDE® (Irinotecan Hydrochloride Liposome), ONTAK® (Denileukin Diftitox), OPDIVO® (Nivolumab), OPPA, Osimertinib, Oxaliplatin, Paclitaxel, Paclitaxel Albumin-stabilized Nanoparticle Formulation, PAD, Palbociclib, Palifermin, Palonosetron Hydrochloride, Palonosetron Hydrochloride and Netupitant, Pamidronate Disodium, Panitumumab, Panobinostat, PARAPLAT® (Carboplatin), PARAPLATIN® (Carboplatin), Pazopanib Hydrochloride, PCV, PEB, Pegaspargase, Pegfilgrastim, Peginterferon Alfa-2b, PEG-INTRON® (Peginterferon Alfa-2b), Pembrolizumab, Pemetrexed Disodium, PERJETA® (Pertuzumab), Pertuzumab, PLATINOL® (Cisplatin), PLATINOL-AQ® (Cisplatin), Plerixafor, Pomalidomide, POMALYST® (Pomalidomide), Ponatinib Hydrochloride, PORTRAZZA® (Necitumumab), Pralatrexate, Prednisone, Procarbazine Hydrochloride, PROLEUKIN® (Aldesleukin), PROLIA® (Denosumab), PROMACTA® (Eltrombopag Olamine), Propranolol Hydrochloride, PROVENGE® (Sipuleucel-T), PURINETHOL® (MMercaptopurine), PURIXAN® (MMercaptopurine), Radium 223 Dichloride, Raloxifene Hydrochloride, Ramucirumab, Rasburicase, R-CHOP, R-CVP, Recombinant Human Papillomavirus (HPV) Bivalent Vaccine, Recombinant Human Papillomavirus (HPV) Nonaval ent Vaccine, Recombinant Human Papillomavirus (HPV) Quadrivalent V accine, Recombinant Interferon Alfa- 2b, Regorafenib, RELISTOR® (Methylnaltrexone Bromide), R-EPOCH, REVLIMID® (Lenalidomide), RHEUM ATREX® (Methotrexate), Ribociclib, R-ICE, RITUXAN® (Rituximab), RITUXAN HYCELA® (Rituximab and Hyaluronidase Human), Rituximab, Rituximab and, Hyaluronidase Human, Rolapitant Hydrochloride, Romidepsin, Romiplostim, RUBIDOMYCIN® (Daunorubicin Hydrochloride), RUBRACA® (Rucaparib Camsylate), Rucaparib Camsylate, Ruxolitinib Phosphate, RYDAPT® (Midostaurin), Sclerosol Intrapleural Aerosol (Talc), Siltuximab, Sipuleucel-T, SOMATULINE DEPOT® (Lanreotide Acetate), Sonidegib, Sorafenib Tosylate, SPRYCEL® (Dasatinib), STANFORD V, Sterile Talc Powder (Talc), STERITALC® (Talc), STIVARGA® (Regorafenib), Sunitinib Malate, SUTENT® (Sunitinib Malate), SYLATRON® (Peginterferon Alfa-2b), SYLVANT® (Siltuximab), Synribo SYNRIBO® (Omacetaxine Mepesuccinate), TABLOID® (Thioguanine), TAG, TAFINLAR® (Dabrafenib), TAGRISSO® (Osimertinib), Talc, Talimogene Laherparepvec, Tamoxifen Citrate,T ARABINE PFS® (Cytarabine), TARCEVA® (Erlotinib Hydrochloride), TARGRETIN® (Bexarotene), TASIGNA® (Nilotinib), TAXOL® (Paclitaxel), TAXOTERE® (Docetaxel), TECENTRIQ® (Atezolizumab), TEMODAR® (Temozolomide), Temozolomide, Temsirolimus, Thalidomide, THALOMID® (Thalidomide), Thioguanine, Thiotepa, Tisagenlecleucel, TOLAK® (Fluorouracil— Topical), Topotecan Hydrochloride, Toremifene, TORISEL® (Temsirolimus), Tositumomab and Iodine I 131 Tositumomab, TOTECT® (Dexrazoxane Hydrochloride), TPF, Trabectedin, Trastuzumab, TREANDA® (Bendamustine Hydrochloride), Trifluridine and Tipiracil Hydrochloride, TRISENOX® (Arsenic Trioxide), TYKERB® (Lapatinib Ditosylate), UNITUXIN® (Dinutuximab), Uridine Tri cetate, VAC, Vandetanib, VAMP, VARUBI® (Rolapitant Hydrochloride), VECTIBIX® (Panitumumab), VelP, VELBAN® (Vinblastine Sulfate), VELCADE® (Bortezomib), VELSAR® (Vinblastine Sulfate), Vemurafenib, VENCLEXTA® (Venetoclax), Venetoclax, VERZENIO® (Abemaciclib), VIADUR® (Leuprolide Acetate), VID AZA® (Azacitidine), Vinblastine Sulfate, VINCASAR PFS® (Vincristine Sulfate), Vincristine Sulfate, Vincristine Sulfate Liposome, Vinorelbine Tartrate, VIP, Vismodegib, VISTOGARD® (Uridine Triacetate), VORAXAZE® (Glucarpidase), Vorinostat, VOTRIENT® (Pazopanib Hydrochloride), VYXEOS® (Daunorubicin Hydrochloride and Cytarabine Liposome), WELLCOVORIN® (Leucovorin Calcium), XALKORI® (Crizotinib), XELODA® (Capecitabine), XELIRI, XELOX, XGEVA® (Denosumab), XOFIGO® (Radium 223 Dichloride), XT ANDI® (Enzalutamide), Y / ERVOY® (Ipilimumab), YONDELIS® (Trabectedin), ZALTRAP® (Ziv-Aflibercept), ZARXIO® (Filgrastim), ZEJULA® (Niraparib Tosyl ate Monohydrate), ZELBORAF® (Vemurafenib), ZEVALIN® ( Ibritumomab Tiuxetan), ZINECARD® (Dexrazoxane Hydrochloride), Ziv-Aflibercept, ZOFRAN® (Ondansetron Hydrochloride), ZOLADEX® (Goserelin Acetate), Zoledronic Acid, ZOLINZ A® (Vorinostat), ZOMETA® (Zoledronic Acid), ZYDELIG® (Idelalisib), ZYKADIA® (Ceritinib), and / or ZYTIGA® (Abiraterone Acetate).

[0167] The treatment methods can include or further include checkpoint inhibitors including, but are not limited to antibodies that block PD-1 (such as, for example, Nivolumab (BMS-936558 or MDX1106), pembrolizumab, cemiplimab, CT-011, MK-3475), PD-L1 (such as, for example, atezolizumab, avelumab, durvalumab, MDX- 1105 (BMS-936559), MPDL3280A, or MSB0010718C), PD-L2 (such as, for example, rHIgM12B7), CTLA-4 (such as, for example, Ipilimumab (MDX-010), Tremelimumab (CP- 675,206)), IDO, B7-H3 (such as, for example,MGA271, MGD009, omburtamab), B7-H4, B7- H3, T cell immunoreceptor with Ig and TTIM domains (TIGIT) (such as, for example BMS- 986207, OMP-313M32, MK-7684, AB-154, ASP-8374, MTIG7192A, or PVSRIPO), CD96, B-and T-lymphocyte attenuator (BTLA), V-domain Ig suppressor of T cell activation (VISTA) (such as, for example, JNJ-61610588, CA-170), TIM3 (such as, for example, TSR-022, MBG453, Sym023, INCAGN2390, LY3321367, BMS-986258, SHR-1702, RO7121661), LAG-3 (such as, for example, BMS-986016, LAG525, MK-4280, REGN 767, TSR-033, BI754111, Sym022, FS118, MGD013, and Immutep).

[0168] In one embodiment, the MEK inhibitor is binimetinib or a therapeutically active analog thereof. In another embodiment, the MEK inhibitor is trametinib or a therapeutically active analog thereof.

[0169] Representative, non-limiting examples of upstream Shp2 inhibitors include TNO155, RMC- 4630, JAB-3068, RLY-1971, ERAS-601, BBP-398, TNO155 and combinations thereof. In one embodiment, the upstream Shp2 inhibitor is PF-07284892 / ARRY-558 or a therapeutically active analog thereof.

[0170] In one embodiment, at least one therapeutic agent is an anti -cancer agent In another embodiment, the anti -cancer agent is an agent that inhibits the proliferation, growth, life-span and / or metastatic activity of a cancer cell. The anti-cancer agent may be, for example, an immune checkpoint inhibitor.

[0171] In one embodiment, the combination of the pan-RAF inhibitor such as a compound of Formula (I)-(IV), and one or more additional therapeutic agents, including but not limited to those disclosed herein, provides a synergistic effect compared to the administration of either compound / agent alone.V. Methods of Manufacture

[0172] The compounds and compositions disclosed herein may be manufactured using any suitable method known in the art.

[0173] For example, the compounds disclosed herein can be prepared according to synthetic methods well known and appreciated in the art. Suitable reaction conditions for the steps of these reactions are well-known in the art and appropriate substitutions of solvents and co-reagents are within the skill of the art. Likewise, it will be appreciated by those skilled in the art that synthetic intermediates may be isolated and / or purified by various well known techniques as needed ordesired, and that frequently, it will be possible to use various intermediates directly in subsequent synthetic steps with little or no purification. Furthermore, the skilled artisan will appreciate that in some circumstances, the order in which moieties are introduced is not critical. The particular order of steps required to produce the compounds of the present invention is dependent upon the compound being synthesized, the starting compound, and the relative liability of the substituted moieties, as is well appreciated by the skilled chemist. All substituents, unless otherwise indicated, are as previously defined, and all reagents are well-known and appreciated in the art.VI. Kits

[0174] The compounds and / or pharmaceutical compositions disclosed herein may be provided as kits. In one embodiment, a kit is provided that includes a pan-RAF inhibitor, a container, and a package insert or label indicating the administration of the pan-RAF inhibitor for a method disclosed herein, including treating, inhibiting, reducing, and / or preventing melanoma. Optionally, the kit may contain a second therapeutic agent.EXAMPLESExample 1: Animal models of melanoma development

[0175] Genetically engineered mouse models (GEMMs), 77V and 7B, replicate the presence of oncogenic NRAS and BRAF mutations in human nevi and melanomas (FIG. 1 A - IB). TN and In the models disclosed herein, melanocyte-specific oncogene induction was induced by painting the animals with 4-hydroxytamoxifen (4OHT) on postnatal days 1 and 2. The mice were then subjected to a single dose of ultraviolet radiation (UV) on postnatal day 3 which mimics the etiologic role of sunlight in melanoma formation (FIG. 1 C ). Representative photographs of nevi on the skin of 7 V and TB mice (FIG. ID). Neonatal exposure of TN and TB mice to 2.0 standard erythemal doses (SEDs) of ultraviolet B irradiation on post-natal day three significantly accelerated melanoma development, resulting in a rate of nevi-to-melanoma transformation comparable to that observed in humans (~1 in 107cells) (FIGS. IE - IF). These models replicate both the genetic and environmental factors involved in human melanoma progression, making them ideal for testing preventive interventions, as disclosed herein.Example 2: NRAS mutant-specific sensitivity to pan-RAF inhibition supports melanomadependency of RAF signaling.

[0176] FIG. 2 is a scheme illustrating type II RAF inhibitors (pan-RAF inhibitors) limits of paradoxical MARK activation. The melanomagenic potential of NRAS mutants generally correlates with BRAF binding affinity (FIGS. 3 A-3B). FIG. 3B is a graphical summary of BRET50 for different Q61 mutations (R = arginine, K = lysine, L = leucine, H = histidine, and P = proline), where lower BRET50 values indicate increased affinity.

[0177] Human melanoma cell lines carrying a strong (Q61 ) or moderate (Q61H) melanoma-driving NRAS oncogene were treated with a low dose of pan-RAF inhibitor for 1 hour and then downstream M APK> ERK signaling was examined. Consistent with the observation that melanomagenic NRAS mutants exhibit enhanced RAF engagement, the pan-RAF inhibitors tested showed greater MAPK> ERK inhibition in the cell line with a moderate (NRAS Q61H) melanoma-driver. FIGS 3C-3D shows representative immunoblots (FIG. 3C) and graphical summary of densitometry measurements (FIG. 3D) of total and phosphorylated ERK in lysates from human melanoma cell lines treated for 1 hour with vehicle (V, DMSO) or doses of pan-RAF inhibitor: (belvarafenib (B), exarafenib (E), or naporafenib (N)). The NZM46 cell line, which harbors an NRAS Q6I H mutant, is more sensitive to pan-RAF inhibition than the SKMel-119 or NZM46 cell lines driven by NRAS Q6IR. * p <0.05, ANOVA. Each dot represents a biological replicate. This finding underscores the importance of using pan-RAF inhibitors at the earliest stages of melanoma development in which RAF engagement has yet to peak and the ceils may be ore sensitive.Example 3: Impact of transient oral pan-RAF inhibition on NRAS- and BRAF-mutant nevi.

[0178] Consistent with the approach in FIG. 4A, litters of TN mice were painted with 4-hydroxytamoxifen (4OHT) on postnatal days 1 and 2 to induce the expression of strong (Q61R) or moderately (Q61H) melanomagenic NRAS mutants in melanocytes. On postnatal day 3 the animals received 2.0 SEDs of solar-simulated UV irradiation, which leads to the formation of nevi.

[0179] At six weeks of age, after nevi formed, the mice were randomly assigned to the control or treatment arm, and high- resolution photographs were taken under anesthesia to document the number or size of nevi on the torso, tail, ear, and paws. The mice received vehicle(control arm) or pan-RAF inhibitor (treatment arm, exarafenib 30 mg / kg or naporafenib 30 mg / kg) orally once a day for three days (Fig. 4A).

[0180] One or five weeks after completing therapy, the mice were photographed and euthanized. The skin was collected, fixed overnight, and processed into paraffin blocks for histopathology and immunohistochemical analyses.

[0181] The following experimental measures are assessed:

[0182] Nevus imaging'. The size, number, and location of nevi are compared between photographs taken at baseline (8 weeks of age) and at the time of euthanasia. Images are converted to black and white, thresholded, and the number and area of each nevus are determined using the ImageJ Analyze particle function. The relative pigmentation of each nevus us measured using the ImageJ region of interest (ROI) tool Endpoint photographs are combined with histopathology in cases of skin toxicity or dynamic nevi.

[0183] Skin histopathology’. Paraffin embedded, H& E or Fontana-Masson stained, dorsal, ear, and tail skin are assessed by a certified veterinary pathologist. Samples are de-identified and scored for evidence of skin toxicity using a 0-4 scale representing normal, mild, moderate, severe, and extreme disease. Emphasis is placed on the visual (photographic) and histopathologic assessment of toxicities reported in clinical trials of exarafenib, including rash, dermatitis acneiform, and pruritus. Additionally, samples are assessed for skin toxicities attributed to other MAPK inhibitors (e.g., keratoses, cutaneous squamous cell carcinomas (SCCs), folliculitis, panniculitis).

[0184] Immunohistochemistry (IHC): Unstained 5 pm skin sections are co-stained for a melanocyte marker (gplOO) and either cleaved Caspase (cell death), phosphorylated ERK (M / kPK activation), or Ki67 (cell proliferation). Slides are scanned and positivity (Caspase, Ki67) or intensity (pERK) scored using Vectra InForm software.

[0185] From these studies, we found that exarafenib reduced skin pigmentation one-week post-therapy in the NRAS Q61H model (FIGS. 4B-4C). Reductions in skin pigmentation were observed in photographs (FIG. 4B) and via quantification of the frequency of Fontana-Masson (melanin) positive cells in histological sections of the skin (FIG. 4C). At 5 weeks post-treatment, the footpads of animals treated with pan-RAF inhibitor (exarafenib or naporafenib) were photographed (FIG. 4D) and scored for pigmentation by blinded reviewers. Reductions in footpad pigmentation were seen in both the exarafenib- and naporafenib-treated groups ascompared to controls (FIG. 4E). These data show that short-term use of an oral pan-RAF inhibitor can selectively eliminate NRAS mutant melanoma precursors from the skin of mice. Similar results are expected following the topical application of pan-RAF inhibitors in both NRAS and BRAF mutant melanoma mouse models.Example 4: Transient pan-RAF inhibition prevents NRAS- and BRAF-mutant melanoma formation.

[0186] Litters of TN mice were painted with 4-hydroxytamoxifen (4OHT) on postnatal days 1 and 2 to induce the expression of strong (Q61R) or moderately (Q61 H) melanomagenic NRAS mutants in melanocytes. On postnatal day 3, the animals received 2.0 SEDs of solar-simulated UV irradiation, which led to the formation of nevi with the potential for melanoma progression. At six weeks of age, after nevi formed, the mice were randomly assigned to the control or treatment arm (FIG. 5A).

[0187] The mice received vehicle (control arm, (5% DMSO, 5% Cremophor EL in water)) or pan-RAF inhibitor (treatment arm, exarafenib 30 mg / kg or naporafenib 30 mg / kg) orally once a day for three days). The mice were then monitored for spontaneous tumor formation, e.g. for up to 52 weeks, and euthanized upon reaching exclusion criteria based on tumor size (>1.6 cm total diameter), ulceration (>2 mm deep), and body condition (score < 2),

[0188] The following experimental measures were determined:

[0189] Tumor onset. A treatment-blinded reviewer checked the study animals for tumors three times a week. Blinding can be achieved by co-housing treated and untreated mice and assigning random numeric study identifiers. The age at which the first tumor is detected in each mouse is recorded, plotted in a Kaplan-Meier curve, and compared.

[0190] Overall survival'. Animals were euthanized at 52 weeks of age or upon reaching the predetermined exclusion criteria described above. The age of the animals at euthanasia is documented, and a full autopsy is performed.

[0191] Tumor growth rate. ' Once a tumor becomes palpable, it was measured daily using digital calipers. The tumor volume was calculated (volume = 0.5 length x width2), and the growth rate was estimated based on the slope of the linear regression plotted over time

[0192] Tumor burden. Total tumor number was recorded during autopsy. Ongoing measurement of individual tumors can ensure proper enumeration of any melanomas that merge prior to the study endpoint.

[0193] Tumor location'. Melanocytes predominantly reside within mouse hair follicles However, in the ear and tail, they are positioned at the epidermal-dermal junctions, resembling human skin. To investigate the potential impact of melanocyte positioning on exarafenib efficacy, the localization of tumors (head, torso, ear / tail, or paw) can be compared between the control and treatment groups.

[0194] Tumor onset and overall survival is assessed using Kaplan-Meier curves and compared through log-rank tests. The relative risk of melanoma development in each group was assessed using hazard ratios (HRs). Tumor burden and growth rate were plotted and statistically analyzed using appropriate tests based on data distribution For example, the Student's t-test or Mann-Whitney test. Chi-squared tests are employed to examine differences in tumor location between the control and treatment groups.

[0195] The results indicated that transient administration of exarafenib or naporafenib prevents melanoma in treated NRAS Q61H (TN6iH) mice (FIG. 5B). Calculated hazard ratios (HRs) indicate a reduced risk of melanoma formation in TN61Hmice treated with either pan-RAF inhibitor as compared to vehicle (V) In the NRAS Q61R model (TN61R), exarafenib shows modest potential as a melanoma preventative (FIG. 5C). A longer treatment window or alternative formulation can improve efficacy. Similar or improved results are expected following the topical application of pan-RAF inhibitors to the NRAS and BRAF mutant melanoma mouse models.Example 5: Topical pan-RAF inhibition delays the onset of NRAS-mutant melanomas.

[0196] TN611* mice were treated with 4-hydroxytamoxifen (4OHT) to induce melanocytespecific oncogenic NRAS expression on postnatal days 1 and 2. They were then subjected to a non-burning dose of UV on post-natal day 3. Topical treatment of the mice with vehicle (W06) or pan-RAF inhibitor began 3 weeks later (FIG. 6A),

[0197] TN6IRmice were treated as depicted in FIG. 6A. At 3 weeks of age, the tails of the mice were treated topically with vehicle (W06), 1% exarafenib, 1% naporafenib, or 1% avutometinib and harvested for immunoblotting 1 hour later. FIG. 6B shows representative immunoblots using the protein from the treated tails. Quantification of MAPK pathway activityphosphorylated ERK / total ERK was measured from the immunoblots (FIG. 6C). Each dot represents a single mouse. Bars shown mean + SEM.

[0198] TN61Rmice were treated as depicted in FIG. 6A. At 3 weeks of age, the tails of the mice were treated topically with vehicle (W06), 1% exarafenib, 1% naporafenib, or 1% avutometinib and harvested for immunobloting 3 hours later. FIG. 6D shows representative immunoblots using the protein from the treated tails. Quantification of MAPK pathway target, DUSP6, normalized to a-Tubulin (FIG. 6E), was measured from the immunoblots shown in FIG.6D.

[0199] TN'! Rmice were treated as depicted in FIG. 6A. At 3 weeks of age, the mice were treated for five consecutive days with topical vehicle (W06), 0.5-1% exarafenib, or 1-3% dabrafenib. Kaplan-Meier curves show that the risk of melanoma formation is reduced by topical exarafenib, but not dabrafenib in TN^1Rmice. (FIG. 6F).Example 6: Topical exarafenib slows the growth of established NRAS-mutant melanomas.

[0200] Litters of 7 A' mice are painted with 4-hy dioxy tamoxifen (4OHT) on postnatal days 1 and 2 to induce the expression of melanomagenic NRAS mutants in melanocytes. On postnatal day 3, the animals receive 4.5 kJ / m2of UVB radiation, which leads to the formation of nevi with the potential for melanoma progression.

[0201] TN61Rmelanomas (7-8 mm in diameter) were treated topically with vehicle (n = 8), 1% exarafenib (n::::8) or 1% avutometinib (n:::5) once daily for up to 10 days. Change in tumor size was measured daily using digital calipers, where data represent % change in tumor size ± SEM) (FIG. 7A). FIG 7B shows a waterfall plot showing percent change in tumor size from baseline to the animal endpoint for the tumors in FIG. 7A.Example 7: pan-RAF inhibitors and melanoma recurrence

[0202] TN61Rand IB mice are painted with 4OHT, UV-irradiated, and monitored for tumor formation. Once tumors reach 10 mm in any direction, the mice are anesthetized and tumors surgically resected, purposely leaving the adjacent margins. One week later, when the surgical wounds are healed, vehicle or the topical MTD of a pan-RAF inhibitor (e.g. avutometinib, belvarafenib, exarafenib, or naporafenib) is applied to the excision area for five days. Tumor re¬ growth is measured daily with digital calipers for 2 months or until exclusion criteria are met.Mice are euthanized at the study endpoint and autopsied for evidence of macrometastases.Suspicious lesions are removed and processed for histopathological assessment,

[0203] Tumor recurrence rates are expected to be high (>80%58). With 20 mice per group (10 male, 10 female), the power can detect at least a 37% difference (44% vs 80%) in the rate of melanoma recurrence at the midpoint (log-rank, a=0.05).

[0204] At least one topical pan-RAF inhibitor is expected to prevent melanoma recurrence and metastasis in the TN1and / or TB models.

[0205] Additional exemplary embodiments contemplated herein are as follows:

[0206] Embodiment 1. A method of reducing the size of a melanoma precursor lesion in a subject in need thereof, the method comprising: non-systemically administering a pan-RAF inhibitor to the melanoma precursor lesion, optionally to inhibit a MAP kinase pathway; wherein the method does not comprise co-administering a downstream ATP competitive inhibitor of the M AP kinase pathway; and wherein the melanoma precursor lesion has not been diagnosed as melanoma, optionally wherein the melanoma precursor lesion has not been pathologically diagnosed as melanoma.

[0207] Embodiment 2. The method of Embodiment 1, wherein the size of the melanoma precursor lesion is reduced by 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20% or 10%.

[0208] Embodiment 3. The method of Embodiment 1 or 2, wherein the size of the melanoma precursor lesion is reduced by 90%-10%, 80%-10%, 70%-10%, 60%-10%, 50%-l 0%, 40%-10%, 30%-l 0%, 20%-10%, 90%-20%, 80%-20%, 70%-20%, 60%-20%, 50%-20%, 40%-20%, 30%-20%, 90%-30%, 80%-30%, 70%-30%, 60%-30%, 50%-30%, 40%-30%, 90%-40%, 80%-40%, 70%-40%, 60%-40%, or 50%-40%.

[0209] Embodiment 4. A method of inhibiting progression of a melanoma precursor lesion to melanoma in a subject in need thereof, the method comprising: non-systemically administering a pan-RAF inhibitor to the melanoma precursor lesion; wherein the method does not comprise coadministering a downstream ATP competitive inhibitor of the MAP kinase pathway; and wherein the melanoma precursor lesion has not been diagnosed as melanoma, optionally wherein the melanoma precursor lesion has not been pathologically diagnosed as melanoma.

[0210] Embodiment 5. The method of Embodiment 4, wherein inhibiting progression comprises not forming a benign nevus, or not forming a dysplastic news, or inhibiting a dysplastic nevus from developing into melanoma.

[0211] Embodiment 6. The method of Embodiment 4, wherein inhibiting progression inhibits Stage 0 melanoma.

[0212] Embodiment 7. The method of any of Embodiments 1 - 6, wherein reducing the size of the melanoma precursor lesion comprises eliminating the melanoma precursor lesion.

[0213] Embodiment 8. A method of preventing melanoma in a subject in need thereof, the method comprising: non-systemically administering a pan-RAF inhibitor to subject; wherein the method does not comprise co-administering a downstream ATP competitive inhibitor of the MAP kinase pathway; and wherein the subject does not have a current melanoma diagnosis, optionally wherein the subject does not have a pathological diagnosis of melanoma.

[0214] Embodiment 9. The method of any of Embodiments 1 - 8, wherein the subject:(a) was born with large congenital nevi; or (b) has atypical or multiple nevus syndrome.

[0215] Embodiment 10. The method of any of Embodiments 1 - 9, wherein the subject does not have melanoma.

[0216] Embodiment 11. The method of any of Embodiments 1 - 10, wherein the subject has not had a melanoma diagnosis for at least 1, 2, 3, 4, or five years,

[0217] Embodiment 12. The method of any of Embodiments 1 - 11, wherein the subject has never been diagnosed with melanoma.

[0218] Embodiment 13. The method of Embodiments 8 - 12, wherein the pan-RAF inhibitor is administered to one or more melanoma precursor lesions.

[0219] Embodiment 14. The method of any of Embodiments 1 - 13, wherein the pan-RAF inhibitor is administered for no more than 12 weeks, no more than 10 weeks, no more than 8 weeks, 6 weeks, no more than 5 weeks, no more than 4 weeks, no more than 3 weeks, no more than 2 weeks, or no more than 7 days.

[0220] Embodiment 15. The method of any of Embodiments 1 - 14, wherein the pan-RAF inhibitor is administered for 6 weeks, 5 weeks, 4 weeks, 3 weeks, 2 weeks or 7 days.

[0221] Embodiment 16. The method of any of Embodiments 1 - 15, wherein the melanoma precursor lesion has not been biopsied.

[0222] Embodiment 17 The method of any of Embodiments 1 - 16, wherein the melanoma precursor lesion has not been incised or excised

[0223] Embodiment 18. The method of any of Embodiments 1 - 17, wherein the melanoma precursor lesion comprises a cluster of melanocytes.

[0224] Embodiment 19. The method of any of Embodiments 1 - 18, wherein the melanoma precursor lesion is a dysplastic nevus.

[0225] Embodiment 20. The method of any of Embodiments 1 - 19, wherein the melanoma precursor lesion comprises cells having NRAS and / or BRAE mutations.

[0226] Embodiment 21. The method of any of Embodiments 1 -20, wherein the melanoma precursor lesion comprises cells having an NRAS mutation at codon 12, 13, or 61, or a BRAF point mutation at residue 594, 600 or 601.

[0227] Embodiment 22 The method of Embodiment 21, wherein the NRAS mutation is Q61R, Q61K, Q61L, Q61H, orQ61P.

[0228] Embodiment 23 The method of Embodiment 21, wherein the NRAS mutation is G12D, G12V, G12C, G12S, G12Aor G12R.

[0229] Embodiment 24. The method of Embodiment 21, wherein the NRAS mutation is G13D or GBR.

[0230] Embodiment 25. The method of Embodiment 21, wherein the BRAF mutation is V600E, V600K, V600R, V600D, V600M, V600G, K601E, or D594N.

[0231] Embodiment 26. A method for inhibiting recurrence of melanoma cells in a border region remaining after excision of a local melanoma in a subject, the method comprising: non-systemically administering a pan-RAF inhibitor to the border region of the subject following the excision of the local melanoma; wherein the method does not comprise co-administering a downstream ATP competitive inhibitor of the MAP kinase pathway.

[0232] Embodiment 27. The method of Embodiment 26, wherein the local melanoma is a Stage 0, Stage I or Stage II melanoma.

[0233] Embodiment 28. The method of Embodiment 26, wherein the subject has not been rediagnosed with melanoma at the site of the local melanoma excision following the excision and prior to non-systemically administering the pan-RAF inhibitor to the border region.

[0234] Embodiment 29. The method of any of Embodiments 1 -28, wherein the subject has not had a melanoma diagnosis for at least one, two, three, four, or five years.

[0235] Embodiment 30 The method of any of Embodiments 1 - 29, wherein the pan-RAF inhibitor inhibits two or more of A-RAF, B-RAF and C-RAF,

[0236] Embodiment 31. The method of any of Embodiments 1 -30, wherein the pan-RAF inhibitor inhibits both protomers of any RAF dimer

[0237] Embodiment 32. The method of any of Embodiments 1 -31, wherein the subject has not been previously treated with a pan-RAF inhibitor.

[0238] Embodiment 33. The method of any of Embodiments 1 - 32, wherein the subject has been previously treated with a pan-RAF inhibitor.

[0239] Embodiment 34. The method of any of Embodiments 1 - 33, wherein the pan RAF inhibitor is AZ628, avutometinib, belvarafenib, brimarafenib, CCT196969, CCT241161, exarafenib, GNE-9815, IHMT-RAF-128, JZP815, lifirafenib, LSN3074753, naporafenib, pan-RAF kinase inhibitor 1, RAF709, SJ-C1044, TBAP-001, tovorafenib, TAK-632 or a therapeutically effective derivative or analog thereof.

[0240] Embodiment 35 The method of any of Embodiments 1 - 33, wherein the pan-RAF inhibitor is selected from the group of AZ628, belvarafenib, CCT 196969, CCT241161, exarafenib, JZP815, and tovorafenib.

[0241] Embodiment 36. The method of any of Embodiments 1 - 33, wherein the pan-RAF inhibitor is exarafenib or naporafenib,

[0242] Embodiment 37. The method of any of Embodiments 1 - 33, wherein the pan-RAF inhibitor is exarafenib.

[0243] Embodiment 38. The method of any of Embodiments 1 - 33, wherein the pan-RAF inhibitor is avutometinib.

[0244] Embodiment 39. The method of any of Embodiments 1 -38, wherein the method does not comprise systemic administration of a second therapeutic agent for the treatment of cancer.

[0245] Embodiment 40. The method of any of Embodiments 1 - 38, wherein the method does not comprise concurrent systemic administration of a second therapeutic agent for the treatment of m elanoma.

[0246] Embodiment 41. The method of any of Embodiments 1 - 38, wherein the method does not comprise systemic administration of a second therapeutic agent for the treatment of melanoma.

[0247] Embodiment 42 The method of any of Embodiments 1 -41, wherein the method does not comprise co- administering an ATP competitive MEK inhibitor,

[0248] Embodiment 43. The method of any of Embodiments 1 - 42, wherein the method does not comprise co-administering an allosteric MEK inhibitor.

[0249] Embodiment 44. The method of any of Embodiments 1 -43, wherein the method does not comprise co-administering an inhibitor of any step of the MAP kinase pathway downstream of RAF, optionally wherein the method does not comprise co-administering a MEK or ERK inhibitor.

[0250] Embodiment 45. The method of any of Embodiments 1 -43, wherein the method does not comprise co-administering an inhibitor of any step of the MAP kinase pathway.

[0251] Embodiment 46. The method of any of Embodiments 1 - 43, wherein the method does not comprise co-administering an inhibitor of one or more of RAS, MEK, or ERK, optionally inhibitor of one or more of RAS, MEK or ERK pathways.

[0252] Embodiment 47 The method of any of Embodiments 1 - 46, wherein the method does not comprise administering a MEK inhibitor.

[0253] Embodiment 48. The method of any of Embodiments 1 -47, wherein the method does not comprise co-administering a mitochondrial inhibitor.

[0254] Embodiment 49. The method of any of Embodiments 1 - 47, wherein the method does not comprise co-administering a mitochondrial respiration inhibitor.

[0255] Embodiment 50. The method of any of Embodiments 1 - 47, wherein the method does not comprise co-administering an oxidative phosphorylation inhibitor.References

[0256] Miller, A. J., & Mihm Jr, M. C. (2006). Melanoma. New England Journal of Medicine, 355(1), 51-65.

[0257] 1. Anderson AM, Matsumoto M, Saul All, Secrest AM, Ferris LK. Accuracy of Skin Cancer Diagnosis by Physician Assistants Compared With Dermatologists in a Large Health Care System. JAMA Dermatol. 2018; 154(5): 569-73. doi: 10.1001 / jamadermatol.2018.0212. PubMed PMID: 29710082; PMCID: PMC6128496.

[0258] 2. Waldmann A, Nolte S. Geller AC, Katalinic A, Weinstock MA, Volkmer B, Greinert R, Breitbart EW. Frequency of excisions and yields of malignant skin tumors in apopulation-based screening intervention of 360,288 whole-body examinations. Arch Dermatol.2012; 148(8) 903- 10. doi: 10.1001 / archdermatol.2012.893. PubMed PMID. 22911184.

[0259] 3. Nault A, Zhang C, Kim K, Saha S, Bennett DD, Xu YG. Biopsy Use in Skin Cancer Diagnosis: Comparing Dermatology Physicians and Advanced Practice Professionals JAMA Dermatol. 2015; 151 (8):899-902. doi: 10.1001 / jamadermatol.2015.0173. PubMed PMID: 25806897.

[0260] 4. Munjal A, Ferguson N. Skin Cancer in Skin of Color. Dermatol Clin.2023;41 (3):481 -9. Epub 20230407. doi: 10.1016 / j.det.2023.02.013. PubMed PMID: 37236716.

[0261] 5. Henrikson NB, Ivlev I, Blasi PR, Nguyen MB, Senger CA, Perdue LA, Lin JS. Screening for Skin Cancer: An Evidence Update for the US Preventive Sendees Task Force. Rockville (MD)2023.

[0262] 6 Kamposioras K, Pentheroudakis G, Pectasides D, Pavlidis N. Malignant melanoma of unknown primary site. To make the long story short. A systematic review of the literature. Critical Reviews in Oncology / Hematology. 2011;78(2):112-26. doi:http s: / / doi. org / 10.1016 / j critrevonc.2010.04.007.

[0263] 7. Helvind NM, Brinch-Moller Weitemeyer M, Chakera AH, Hendel HW, Ellebaek E, Svane IM, Kjaerskov MW, Bojesen S, Skyum H, Petersen SK, Bastholt L, Johansen C, Bidstrup PE, Holmich LR. Stage-Specific Risk of Recurrence and Death From Melanoma in Denmark, 2008-2021: A National Observational Cohort Study of 25720 Patients With Stage IA to IV Melanoma. JAMA Dermatol. 2023; 159(11): 1213-22. doi:10.1001 / jamadermatol.2023.3256. PubMed PMID: 37650576; PMCID: PMC10472263.

[0264] 8. Castilla EE, da Graca Dutra M, Orioli-Parreiras IM. Epidemiology of congenital pigmented naevi: I. Incidence rates and relative frequencies. Br J Dermatol. 1981; 104(3):307-l 5. doi: 10.1111 / j.1365-2133.1981.tb00954.x. PubMed PMID: 7213564.

[0265] 9. Tromberg J, Bauer B, Benvenuto-Andrade C, Marghoob AA. Congenital melanocytic nevi needing treatment. Dermatol Ther. 2005; 18(2): 136-50. doi: 10.1111 / j.1529-8019.2005.05012.x. PubMed PMID: 15953143.

[0266] 10 Gu J, Wei B, Duan R, Jin J, Lai B, Wei J, Xie F. Insight into the Relationship between Congenital Large / Giant Melanocytic Nevi and Satellite Nevi. J Invest Dermatol. 2025. Epub 20250129. doi: 10.1016 / j jid.2024.10.620 PubMed PMID: 39890054.

[0267] 11. MacKie RM, Freudenberger T, Aitchison TC. Personal risk-factor chart for cutaneous melanoma. Lancet, 1989,2(8661 ):487-90. doi: 10,1016 / s0140-6736(89)92097-7. PubMed PMID: 2570195.

[0268] 12. SEER*Explorer: An interactive website for SEER cancer statistics [Internet], 2024. Available from: https: / / seer.cancer.gov / statistics-network / explorer / .

[0269] 13. Zeng H, Judson-Torres RL, Shain AH. The Evolution of Melanoma - Moving beyond Binaiy Models of Genetic Progression. J Invest Dermatol. 2020;140(2):291-7. Epub 20191014. doi: 10.1016 / j.jid.2019.08.002. PubMed PMID: 31623932; PMCID: PMC6983335.

[0270] 14. Shain AH, Joseph NM, Yu R, Benhamida J, Liu S, Prow T, Ruben B, North J, Pincus L, Yeh I, Judson R, Bastian BC. Genomic and Transcriptomic Analysis Reveals Incremental Disruption of Key Signaling Pathways during Melanoma Evolution. Cancer Cell. 2018;34(l):45-55 e4. doi: 10.1016 / j ccell.2018.06.005 PubMed PMID: 29990500: PMCID: PMC6319271.

[0271] 15. Yeh I, von Deimling A, Bastian BC. Clonal BRAF mutations in melanocytic nevi and initiating role of BRAF in melanocytic neoplasia. J Natl Cancer Inst. 2013, 105(12):917-9. Epub 20130520. doi: 10.1093 / jnci / djtl 19. PubMed PMID: 23690527; PMCID: PMC3687371.

[0272] 16. Maldonado JL, Fridlyand J, Patel H, Jain AN, Busam K, Kageshita T, Ono T, Albertson DG, Pinkel D, Bastian BC. Determinants of BR AF mutations in primary melanomas. J Natl Cancer Inst. 2003;95(24): 1878-90. doi: 10.1093 / jnci / djgl23. PubMedPMID: 14679157.

[0273] 17. Busam KJ, Hedvat C, Pulitzer M, von Deimling A, Jungbluth AA Immunohistochemical analysis of BRAF(V600E) expression of primary and metastatic melanoma and comparison with mutation status and melanocyte differentiation antigens of metastatic lesions. Am J Sur Pathol. 2013;37(3 ):413-20. doi: 10.1097 / PAS.0b013e318271249e. PubMedPMID: 23211290.

[0274] 18. Burd CT.. Liu W, Huynh MV, Waqas MA, Gillahan JE, Clark KS, Fu K, Martin BL, Jeck WR, Souroullas GP, Darr DB, Zedek DC, Miley MJ, Baguley BC, Campbell SL, Sharpless NE. Mutation-specific RAS oncogenicity explains NRAS codon 61 selection in melanoma. Cancer Discov. 2014;4(12): 1418-29. Epub 20140924. doi: 10.1158 / 2159-8290. CD- 14-0729. PubMed PMID: 25252692; PMCID: PMC4258185.

[0275] 19. Murphy BM, Terrel! EM, Chirasani VR, Weiss TJ, Lew RE, Holderbaum AM, Dhakal A, Posada V, Fort M, Bodnar MS, Carey LM, Chen M, Burd CJ, Coppola V, MorrisonDK, Campbell SL, Burd CE. Enhanced BRAF engagement by NRAS mutants capable of promoting melanoma initiation Nat Commun. 2022; 13(1 ):3153 Epub 20220607. doi:10.1038 / s41467-022-30881-9. PubMedPMID: 35672316; PMCID: PMC9174180.

[0276] 20. Chen FW, Tseng D, Reddy S, Daud Al, Swetter SM. Involution of eruptive melanocytic nevi on combination BRAF and MEK inhibitor therapy. JAMA Dermatol.2014;150(l 1): 1209-12. doi: 10.1001 / jamadermatol.2014.838. PubMedPMID: 25142409.

[0277] 21. Boussemart L, Routier E, Mateus C, Opletalova K, Sebille G, Kamsu-Kom N, Thomas M, Vagner S, Favre M, Tomasi c G, Wechsler J, Lacroix L, Robert C. Prospective study of cutaneous side-effects associated with the BRAF inhibitor vemurafenib: a study of 42 patients. Ann Oncol. 2013;24(6):1691-7. Epub 20130213. doi: 10.1093 / annonc / mdt015. PubMed PMID: 23406731.

[0278] 22. Haenssle HA. Kraus SL. Brehmer F, Kretschmer L, Volker B, Asper H, Kapp A, Gutzmer R. Dynamic changes in nevi of a patient with melanoma treated with vemurafenib: importance of sequential dermoscopy. Arch Dermatol. 2012; 148(10): 1183-5. doi:10.1001 / archdermatol.2012.2649. PubMed PMID: 22911096.

[0279] 23. McClenahan P, Lin LL, Tan IM, Flewell-Smith R, Schaider H, Jagirdar K, Atkinson V, Lambie D, Prow TW, Sturm RA, Soyer HP. BRAFV600E mutation status of involuting and stable nevi in dabrafenib therapy with or without trametinib. JAMA Dermatol. 2014;150(10):1079-82. doi: 10.1001 / jamadermatol.2014.436. PubMedPMID: 24695877.

[0280] 24. Parekh V, Sobanko J, Miller CJ, Karakousis G, Xu W, Letrero R, Elenitsas R, Xu X, Elder DE, Amaravadi R, Schuchter LM, Nathanson KL, Wilson MA, Chu EY. NRAS Q61R and BRAF G466A mutations in atypical melanocytic lesions newly arising in advanced melanoma patients treated with vemurafenib. J Cutan Pathol. 2019;46(3):190-4. Epub 20181227. doi: 10.1111 / cup.13401. PubMed PMID: 30552700; PMCID: PMC6367041.

[0281] 25. Perier-Muzet M, Boespflug A, Poulalhon N, Caramel J. Breton AL, Thomas L, Dalle S. Dermoscopic Evaluation of Melanocytic Nevi Changes With Combined Mitogen- Activated Protein Kinase Pathway Inhibitors Therapy for Melanoma. J AMA Dermatol.2016:152(10): 1162-4. doi: 10.1001 / jamadermatol.2016.2426. PubMedPMID: 27463731.

[0282] 26. Giurcaneanu C, Nitipir C, Popa LG, Forsea AM, Popescu I, Bumbacea RS Evolution of melanocytic nevi under vemurafenib, followed by combination therapy withdabrafenib and trametinib for metastatic, melanoma Acta Dermatovenerol Croat 2015;23(2): 114-21. PubMed PMID: 26228819.

[0283] 27. Murphy BM, Burd CE. Can Combination MEK and Akt Inhibition Slay the Giant Congenital Nevus J Invest Dermatol. 2019; 139(9): 1857-9. doi: 10.1016 / j.jid.2019.04.009. PubMed PMID: 31445573; PMCID: PMC6921933.

[0284] 28. Pawlikowski JS, Brock C, Chen SC, Al-Olabi L, Nixon C, McGregor F, Paine S, Chanudet E, Lambie W, Holmes WM, Mullin JM, Richmond A, Wu H, Blyth K, King A, Kinsler VA, Adams PD. Acute Inhibition of MEK Suppresses Congenital Melanocytic Nevus Syndrome in a Murine Model Driven by Activated NRAS and Wnt Signaling J Invest Dermatol.2015;135(ll):2902. Epub 20150716. doi: 10.1038 / jid.2015.230. PubMed PMID: 26178707.

[0285] 29. Choi YS, Erlich TH, von Franque M, Rachmin I, Flesher JL, Schiferle EB, Zhang Y, Pereira da Silva M, Jiang A, Dobry AS, Su M, Germana S, Lacher S, Freund O, Feder E, Cortez JL, Ryu S, Babila Propp T, Samuels YL, Zakka LR, Azin M, Burd CE, Sharpless NE, Liu XS, Meyer C, Austen WG, Jr., Bojovic B, Cetailo CL, Jr., Mihm MC, Hoon DS, Demehri S. Hawryluk EB, Fisher DE. Topical therapy for regression and melanoma prevention of congenital giant nevi. Cell. 2022;185(12):2071-85 e!2. Epub 20220512. doi: 10.1016 / j.cell.2022.04.025. PubMed PMID: 35561684; PMCID: PMC9237838.

[0286] 30, Poulikakos PI, Zhang C, Bollag G, Shokat KM, Rosen N. RAF inhibitors transactivate RAF dimers and ERK signalling in cells with wild-type BRAF. Nature.2010;464(7287):427-30 doi: 10.1038 / nature08902. PubMed PMID: 20179705; PMCID:PMC3178447.

[0287] 31. Su F, Viros A, Milagre C, Trunzer K, Bollag G, Spleiss O, Reis-Filho JS, Kong X, Koya RC, Flaherty KT, Chapman PB, Kim MJ, Hayward R, Martin M, Yang H, Wang Q, Hilton FI, Hang JS, Noe J, Lambros M, Geyer F, Dhomen N, Niculescu-Duvaz I, Zambon A, Niculescu-Duvaz D, Preece N, Robert L, Otte NJ, Mok S, Kee D, Ma Y„ Zhang C, Habets G, Burton EA, Wong B, Nguyen H, Kockx M, Andries L, Lestini B, Nolop KB, Lee RJ, Joe AK, Troy JL, Gonzalez R, Hutson TE, Puzanov I, Chmielewski B, Springer CJ, McArthur GA, Sosman JA, Lo RS, Ribas A, Marais R. RAS mutations in cutaneous squamous-cell carcinomas in patients treated with BRAF inhibitors. N Engl J Med. 2012;366(3):207-15 doi:10.1056 / NEJMoal 105358. PubMed PMID: 22256804; PMCID: PMC3724537.

[0288] 32. Yen I, Shanahan F, Lee J, Hong YS, Shin SJ, Moore AR, Sudhamsu J, Chang MT, Bae I, Dela Cruz D, Hunsaker T, Klijn C, Liau NPD, Lin E, Martin SE, Modrusan Z, Piskol R, Segal E, Venkatanarayan A, Ye X, Yin J, Zhang L, Kim JS, Lim HS, Kim KP, Kim YJ, Han HS, Lee SJ, Kim ST, Jung M, Hong YH, Noh YS, Choi M, Han O, Nowicka M, Srinivasan S, Yan Y, Kim TW, Malek S. ARAF mutations confer resistance to the RAF inhibitor belvarafenib in melanoma. Nature. 202i;594(7863):418-23. Epub 20210505. doi: 10.1038 / s41586-021 - 03515-1. PubMed PMID: 33953400.

[0289] 33. Cassier P, Mehnert JM, Kim R, Chmielowski B, Millward M, Perez CA, Gonzalez Cao M, Gambardella V, McKean M, Brose MS, Xing Y, Long GV, Kelly A, Lee G, Severson P, Williams R, Gaudy Marqueste C. 613MO A phase I clinical trial evaluating exarafenib, a selective pan-RAF inhibitor in combination with binimetinib in NRAS-mutant (NRASMut) melanoma (Mel) & BRAF-altered solid tumors. Annals of Oncology.2024;35: S491-S2. doi: 10.1016 / j.annonc.2024.08.680.

[0290] 34. Kim TW, Lee J, Kim TM, Shin SJ, Han SW, Kim JS, Kim YJ, Yoo C, Hong YS, Kim JW, Lee DH, Ahn CB, Kim ST, Kim JW, Hong YH, Kim J, Baek E, Choi B, Malhi V, Baek S. 529P A phase lb trial of belvarafenib in combination with cobimetinib in patients (pts) with RAS- or RAF- mutated (m) solid tumors: Updated safety data and indication-specific efficacy results. Annals of Oncology. 2021;32: S595. doi: 10.1016 / j.annonc.2021.08.1051.

[0291] 35. Hennessey RC, Holderbaum AM, Bonilla A, Delaney C, Gillahan JE, Tober KL, Oberyszyn TM, Zippin JH, Burd CE. Ultraviolet radiation accelerates NRas-mutant melanomagenesis: A cooperative effect blocked by sunscreen. Pigment Cell Melanoma Res. 2017;30(5):477-87. Epub 20170704. doi: 10.1111 / pcmr.12601. PubMed PMID: 28544727.

[0292] 36. The Cancer Genome Atlas Network. Genomic Classification of Cutaneous Melanoma. Cell. 2015;161(7): 1681-96. doi: 10.1016 / j.cell.2015.05.044. PubMed PMID:26091043; PMCID: PMC4580370.

[0293] 37. Shain AH, Yeh I, Kovalyshyn I, Sriharan A, Talevich E, Gagnon A, Dummer R, North J, Pincus L, Ruben B, Rickaby W, D'Arrigo C, Robson A, Bastian BC. The Genetic Evolution of Melanoma from Precursor Lesions. N Engl J Med. 2015;373 (20): 1926-36. doi: 10.1056, / NEJMoal502583. PubMed PMID: 26559571.

[0294] 38. Bauer J, Curtin JA, Pinkel D, Bastian BC Congenital melanocytic nevi frequently harbor NRAS mutations but no BRAF mutations. J Invest Dermatol. 2007; 127(1): 179-82, Epub 20060803. doi: 10.1038 / sj jid.5700490. PubMed PMID: 16888631.

[0295] 39. Melamed RD, Aydin IT, Rajan GS, Phelps R, Silvers DN, Emmett KJ, Brunner G, Rabadan R, Celebi JT. Genomic Characterization of Dysplastic Nevi Unveils Implications for Diagnosis of Melanoma. J Invest Dermatol. 2017;137(4):905-9. Epub 20161124. doi:10.1016 / j jid.2016.11.017. PubMed PMID: 27890785.

[0296] 40. Roh MR, Eliades P, Gupta S. Tsao H. Genetics of melanocytic nevi. Pigment Cell MelanomaRes. 2015;28(6):661-72. doi: 10.111 l / pcmr 12412. PubMed PMID: 26300491, PMCID: PMC4609613.

[0297] 41. Tsao H, Bevona C, Goggins W, Quinn T. The transformation rate of moles (melanocytic nevi) into cutaneous melanoma: a population-based estimate Arch Dermatol. 2003;139(3):282-8. doi: 10.1001 / archderm.139.3.282. PubMed PMID: 12622618.

[0298] 42. Hirobe T, Takeuchi T. Induction of melanogenesis in the epidermal melanoblasts of newborn mouse skin by MSH. J Embryol Exp Morphol. 1977;37(l):79-90. PubMed PMID: 192826.

[0299] 43. Bosenberg M, Muthusamy V, Curley DP, Wang Z, Hobbs C, Nelson B, Nogueira C, Horner JW, 2nd, Depinho R, Chin L, Characterization of melanocyte-specific inducible Cre recombinase transgenic mice. Genesis. 2006;44(5):262-7. doi: 10.1002 / dvg.20205. PubMed PMID: 16676322.

[0300] 44. Dankort D, Curley DP, Cartlidge RA, Nelson B, Kamezis AN, Damsky WE, Jr., You MJ, DePinho RA, McMahon M, Bosenberg M. Braf(V600E) cooperates with Pten loss to induce metastatic melanoma. Nat Genet. 2009;41(5):544-52. Epub 20090312. doi:10.1038 / ng.356. PubMed PMID: 19282848; PMCID: PMC2705918.

[0301] 45. Monahan KB, Rozenberg GI, Krishnaraurthy J, Johnson SM, Liu W, Bradford MK, Horner J, Depinho RA, Sharpless NE. Somatic pl6(INK4a) loss accelerates melanomagenesis. Oncogene. 20I0;29(43):5809-l 7. Epub 20100809, doi:10.1038 / onc.2010.314. PubMed PMID: 20697345; PMCID: PMC3007178.

[0302] 46. Wang TS, Lee C, Severson P, Pelham RJ, Williams R, Miller NLG. Abstract 4927: Exarafenib (KIN-2787) is a potent, selective pan-RAF inhibitor with activity in preclinicalmodels of BRAF class II / ITI mutant and NRAS mutant melanoma. Cancer Research. 2023;83(7__Supplement):4927~. doi: 10.1158 / 1538-7445. Am2023-4927.

[0303] 47. Goetz A, Shanahan F, Brooks L, Lin E, Mroue R, Dela Cruz D, Hunsaker T, Czech B, Dixit P, Segal U, Martin S, Foster SA, Gerosa L. Computational Modeling of Drug Response Identifies Mutant-Specific Constraints for Dosing panRAF and MEK Inhibitors in Melanoma. Cancers (Basel). 2024; 16(16). Epub 20240822. doi: 10.3390 / cancersl6162914. PubMedPMID: 39199684; PMCID: PMC11353013.

[0304] 48. Chen YK, Kanouni T, Arnold LD, Cox JM, Gardiner E, Grandinetti K, Jiang P, Kai dor SW, Lee C, Li C, Martin ES, Miller N, Murphy EA, Timple N, Tyhonas JS, Vassar A, Wang TS, Williams R, Yuan D, Kania RS. The Discovery of Exarafenib (KIN-2787):Overcoming the Challenges of Pan-RAF Kinase Inhibition. J Med Chem. 2024,67(3): 1747-57, Epub 20240117. doi: 10 1021 / acs.jmedchem.3c01830 PubMed PMID: 38230963.

[0305] 49. Cheung MC, Spalding PB, Gutierrez JC, Balkan W, Namias N, Koniaris LG, Zimmers TA. Body Surface Area Prediction in Normal, Hypermuscular, and Obese Mice.Journal of Surgical Research. 2009;153(2):326- 1. doi: https: / / doi. Org / 10.1016 / j.jss.2008.05.002.

[0306] 50. Long CC, Finlay AY, Averill RW. The rule of hand: 4 hand areas = 2 FTU = 1 g. Arch Dermatol. 1992; 128(8): 1129-30. PubMed PMID: 1497374.

[0307] 51. Lawrence J, Seelig D, Demos-Davies K, Ferreira C, Ren Y, Wang L, Alam SK, Yang R, Guedes A, Craig A, Hoeppner LH. Radiation dermatitis in the hairless mouse model mimics human radiation dermatitis. Sci Rep. 2024; 14(1 ):24819. Epub 20241022 doi:10.1038 / s41598-024-76021-9. PubMed PMID: 39438583; PMCID: PMC11496547.

[0308] 52. Belote RL, Le D, Maynard A, Lang UE, Sinclair A, Lohman BK, Planells-Palop V, Baskin L, Tward AD, Darmanis S, Judson-Torres RL. Human melanocyte development and melanoma dedifferentiation at single-cell resolution. Nat Cell Biol. 2021;23(9): 1035-47. Epub 20210902. doi: 10 1038 / s41556-021-00740-8 PubMed PMID: 34475532.

[0309] 53. Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA, Paulovich A, Pomeroy SL, Golub TR, Lander ES, Mesirov JP. Gene set enrichment analysis. A knowledge-based approach for interpreting genome-wide expression profiles. Proceedings of the National Academy of Sciences. 2005;102(43): 15545-50. doi: doi: 10.1073 / pnas.0506580102.

[0310] 54. Dann E, Henderson NC, Teichmann SA, Morgan MD, Marioni JC. Differential abundance testing on single-cell data using k-nearest neighbor graphs. Nat Biotechnol.2022;40(2):245-53. Epub 20210930. doi: 10.1038 / s41587-021 -01033-z PubMed PMID:34594043; PMCID: PMC7617075.

[0311] 55. Zhao J, Jaffe A, Li H, Lindenbaum O, Sefik E, Jackson R, Cheng X, Flavell RA, Kluger Y Detection of differentially abundant cell subpopulations in scRNA-seq data. Proc Natl Acad Sci U S A. 2021;118(22). doi: 10.1073 / pnas.2100293118. PubMed PMID: 34001664; PMCID: PMC8179149.

[0312] 56. Vasta JD, Michaud A, Zimprich CA, Beck MT, Swiatnicki MR, Zegzouti H, Thomas MR, Wilkinson J, Crapster JA, Robers MB. Protomer selectivity of type II RAF inhibitors within the RAS / RAF complex. Cell Chemical Biology. 2023;30( 11 ): 1354-65 e6. doi: https: / / doi.org / 10.1016 / j.chembiol.2023.07.019.

[0313] 57. Monaco KA, Delach S, Yuan J, Mishina Y, Fordjour P, Labrot E, McKay D, Guo R, Higgins S, Wang HQ, Liang J, Bui K, Green J, Aspesi P, Ambrose J, Mapa F, Griner L, Jaskelioff M, Fuller J, Crawford K, Pardee G, Widger S, Hammerman PS, Engelman JA, Stuart DD, Cooke VG, Caponigro G. LXH254, a Potent and Selective ARAF-Sparing Inhibitor of BRAF and CRAF for the Treatment of MAPK -Driven Tumors. Clin Cancer Res.2021;27(7):2061-73. Epub 20201222. doi: 10.1158 / 1078-0432. CCR-20-2563. PubMed PMID: 33355204.

[0314] 58, Lobos-Gonzalez. L, Aguilar-Guzman L, Fernandez JG, Munoz N, Hossain M, Bieneck S, Silva V, Burzio V, Sviderskaya EV, Bennett DC, Leyton L, Quest AF. Caveolin-1 is a risk factor for postsurgery metastasis in preclinical melanoma models. Melanoma Res.2014;24(2):108-19. doi: 10.1097 / CMR.0000000000000046. PubMedPMID: 24500501;PMCID: PMC3999137.

[0315] 59. Yin H, Tang Q, Xia H, Bi F. Targeting RAF dimers in RAS mutant tumors: From biology to clinic. Acta Pharm Sin B. 2024; 14(5): 1895-923. Epub 20240228. doi:10.1016 / j.apsb.2024.02.018. PubMed PMID: 38799634; PMCID: PMC11120325.

[0316] 60. Sarkisian S, Davar D. MEK inhibitors for the treatment of NRAS mutant melanoma. Drug Des Devel Ther. 2018;12:2553-65, Epub 20180820. doi:10.2147 / DDDT. S 131721. PubMed PMID: 30154648; PMCID: PMC6108333.

[0317] All patent documents referred to herein are incorporated by reference in their entireties. Various embodiments of the invention have been described in fulfillment of thevarious objectives of the invention. It should be recognized that these embodiments are merely illustrative of the principles of the present invention. Numerous modifications and adaptations thereof will be readily apparent to those skilled in the art without departing from the spirit and scope of the invention.

Claims

CLAIMS1. A method of reducing the size of a melanoma precursor lesion in a subject in need thereof, the method comprising:non-systemically administering a pan-RAF inhibitor to the melanoma precursor lesion; wherein the method does not comprise co-administering a downstream ATP competitive inhibitor of the MAP kinase pathway; andwherein the melanoma precursor lesion has not been diagnosed as melanoma.

2. A method of inhibiting progression of a melanoma precursor lesion to melanoma in a subject in need thereof, the method comprising:non-systemically administering a pan-RAF inhibitor to the melanoma precursor lesion; wherein the method does not comprise co-administering a downstream ATP competitive inhibitor of the M AP kinase pathway, andwherein the melanoma precursor lesion has not been diagnosed as melanoma.

3. A method of preventing melanoma in a subject in need thereof, the method comprising:non-systemically administering a pan-RAF inhibitor to the subject;wherein the method does not comprise co-administering a downstream ATP competitive inhibitor of the MAP kinase pathway; andwherein the subject does not have a current melanoma diagnosis.

4. The method of any of claim 1, 2 or 3, wherein the subject has never been diagnosed with melanoma.

5. The method of any of claim 1, 2, or 3, wherein the subject:(a) was born with large congenital nevi; or(b) has atypical or multiple nevus syndrome.

6. The method of any of claim 1, 2, or 3, wherein the pan-RAF inhibitor is administered for fewer than 4 weeks.

7. The method of claim 1 or 2, wherein the melanoma precursor lesion has not been biopsied, incised, or excised.

8. The method of claim 1 or 2, wherein the melanoma precursor lesion comprises a cluster of melanocytes.

9. The method of claim 1 or 2, wherein the melanoma precursor lesion is a dysplastic nevus.

10. The method of claim 1 or 2, wherein the melanoma precursor lesion comprises cells having an NRAS mutation at codon 12, 13, or 61, or a BRAF point mutation at residue 594, 600 or 601.

11. The method of claim 10, wherein the NRAS mutation is Q61R, Q61K, Q61L, Q61H, or Q61P.

12. The method of claim 10, wherein the NRAS mutation is G12D, GI2V, G12C, G12S, G12A or G12R.

13. The method of claim 10, wherein the NRAS mutation is G13D or G13R.

14. The method of claim 10, wherein the BRAF mutation is V600E, V600K, V600R, V600D, V600M, V600G, K601E, or D594N.

15. A method for inhibiting recurrence of melanoma in a border region remaining after excision of a local melanoma in a subject, the method comprising:non-systemically administering a pan-RAF inhibitor to the border region of the subject following the excision of the local melanoma;wherein the method does not comprise co-administering a downstream ATP competitive inhibitor of the MAP kinase pathway.16 The method of claim 15, wherein the local melanoma is a Stage 0, Stage I or Stage II melanoma.

17. The method of claim 15, wherein the subject has not been re-diagnosed with melanoma at the site of the local melanoma excision following the excision and prior to non-systemically administering the pan-RAF inhibitor to the border region.18 The method of any of claims I, 2, 3 or 15, wherein the pan RAF inhibitor is AZ628, avutometinib, belvarafenib, brimarafenib, CCT 196969, CCT241161, exarafenib, GNE-9815, IHMT-RAF-128, JZP81538, lifirafenib, LSN3074753, naporafenib, pan-RAF kinase inhibitor 1, RAF709, SJ-C1044, TBAP-001, tovorafenib, TAK-632 or a therapeutically effective derivative or analog thereof,19. The method of any of claims 1, 2, 3 or 15, wherein the method does not comprise co¬ administering an ATP competitive MEK inhibitor.20 The method of any of claims 1, 2, 3 or 15, wherein the method does not comprise coadministering an allosteric MEK inhibitor.

21. The method of any of claims 1, 2, 3 or 15, wherein the method does not comprise coadministering an inhibitor of the MAP kinase pathway.

Images