AU2021360676B2

AU2021360676B2 - Composition and method for treating cancer using a vaccine as a first therapueutic active ingredient in combination with a second active ingredient

Info

Publication number: AU2021360676B2
Application number: AU2021360676A
Authority: AU
Inventors: Evangelos V. BADIAVAS; Tim Ioannides
Original assignee: Hpvvax LLC
Current assignee: Hpvvax LLC
Priority date: 2020-10-12
Filing date: 2021-10-12
Publication date: 2025-09-04
Anticipated expiration: 2041-10-12
Also published as: HUE21880926T1; EP4225351A1; PL4225351T1; EP4225351A4; CA3195478A1; CO2023006083A2; MX2023004285A; IL302098A; KR20230117106A; WO2022081604A1; JP2023546073A; DK4225351T1; ES2962700T1; DE21880926T1; AU2021360676A9; AU2021360676A1; CY20232200001T2; FI4225351T1

Abstract

A method for treating or reducing the incidence of recurrence of cancer, benign tumors, or HPV-associated lesions, including skin cancer, and particularly squamous cell carcinoma (SCC and basal-cell carcinoma, by administering to a patient one or more doses of HPV recombinant vaccine as a first active therapeutic agent in combination with a second active therapeutic agent administered concomitantly or as a fixed-dose combination composition.

Description

WO wo 2022/081604 PCT/US2021/054622 PCT/US2021/054622

COMPOSITION AND METHOD FOR TREATING CANCER USING A VACCINE AS A FIRST THERAPUEUTIC ACTIVE INGREDIENT IN COMBINATION WITH A SECOND ACTIVE INGREDIENT FIELD OF THE INVENTION

[0001] The invention relates to treating cancer, including skin cancer or malignant

tumor and, more particularly, to a composition and method for treatment, or reducing the

incidence or recurrence of, cancer or tumor by administration of a vaccine such as a

conventional human papillomavirus (HPV) vaccine, as a first active therapeutic agent in

combination with a second active therapeutic agent administered concomitantly or as a fixed-

dose combination composition.

BACKGROUND OF THE INVENTION

[0002] Skin cancer consists of three main types, namely, basal-cell carcinoma (BCC),

squamous cell carcinoma (SCC) and melanoma. Skin cancer is the most common form of

cancer globally. Understandably, there have been ongoing studies for many years searching

for effective methods to treat, and possibly cure, these types of skin cancer.

[0003] It is generally accepted that human papillomavirus (HPV) is associated with

causing certain types of skin cancer, particularly squamous cell carcinoma (SCC). HPV is a

DNA virus that can infect certain types of tissues in humans. There are upwards of thirty

subtypes of HPV and some of these subtypes have been associated with cervical cancer,

including HPV16 and HPV18. HPV is not known to be a cause or to be associated with basal

cell carcinoma (BCC) or melanoma. Basal-cell carcinoma (BCC) and melanoma are generally accepted as being unrelated to or independent from HPV infection.

[0004] Vaccines have been developed and shown to prevent cervical cancer in women

and other conditions caused by or associated with HPV infection. GARDASIL® is a

commercially available vaccine having activity against HPV (types 6, 11, 16, and 18).

[0005] GARDASIL® 9 is another commercially available vaccine marketed for prevention of HPV (types 16, 18, 31, 33, 45, 52, and 58). GARDASIL® is indicated GARDASIL is indicated for for use use

in girls and boys from ages 9-26; GARDASIL® GARDASIL 99 is is also also indicated indicated for for use use in in girls girls from from ages ages

9-26, and in boys from ages 9-15.

PCT/US2021/054622

[0006] Other vaccines have been produced, as well, for treating subtypes of HPV,

particularly HPV16 and HPV18. GARDASIL® and other known vaccines administered

prophylactically, prophylactically, to to prevent prevent certain certain HPV HPV infections infections and and associated associated cancers, cancers, are are referred referred to to

herein as "preventive vaccines." These preventive vaccines are typically administered for

systemic action, being injected into a patient subcutaneously or intramuscularly (e.g.,

deltoid), remote from any particular target, such as the cervix. Moreover, they are generally

accepted to be effective prior to exposure to HPV and are not commonly known to be

effective for treatment after exposure to, or infection with, HPV.

[0007] Other preventive vaccines include, for example, an improved vaccine

composition as described in Chinese Pat App. No. 101890160 (CN'160) comprising certain

L1 L1 proteins proteinsofofHPV (as(as HPV in in GARDASIL ), and and GARDASIL), additional HPV-specific additional components. HPV-specific components.

Preventive vaccines comprising HPV-type 16 and 18 proteins are also suggested to provide

cross-protection against other HPV types, as described in US Pub. No. 2005/0287161.

[0008] Vaccines used for treatment (referred to herein as "therapeutic vaccines") are

described. However, these therapeutic vaccines are understood in the art to require more than

viral-specific components, such as the HPV L1 proteins that comprise the commercially

available preventive vaccines, such as GARDASIL®. GARDASIL®

[0009] US Pub. No. 2007/0218074 describes the use of a vaccine composition

comprising host-cell peptides from an HPV-infected cell. The host-cell peptides, e.g., the

early antigens, E6 or E7, that present on the surface of cells infected with HPV, are fragments

of host-cell proteins. The criticality of the polypeptides E6 or E7 in a vaccine used in treating

certain cancer types is described in Development of HPV vaccines for HPV-associated head

and neck squamous cell Carcinoma, Devaraj, et al., Crit Rev Oral Biol Med. 2003;14(5):345-

62. Another vaccine which includes a host-cell protein (BAX) is described in US Pat. No.

8,399,610.

[00010] Yet another vaccine composition comprising other or additional antigens in

combination with HPV-16 peptides, is a vaccine composition described in US Pub. No.

2011//0070252 which additionally 2011/0070252 which additionally requires requires Trojan Trojan antigen. antigen.

[00011] US Pub. No. 2011/0110979 (US '979) and US Pub. No. 2012/0288538 (US

'538) disclose therapeutic use of an HPV vaccine comprising E6 or E7 polypeptides (peptide fragments from host cells infected with HPV). US '538 describes that E6 and E7 are crucial to induce transformation into HPV-infected cells, and states that a vaccine composition which does not include E6 or E7 would not be expected to work on cells that do not have E6 or E7, i.e., cells such as BCC that are not infected with HPV. The method described in the US'979 publication additionally requires an immunostimulant or adjuvant.

[00012] Although the US '979 and US '538 publications describe the use of therapeutic

vaccines against skin cancers, such as SCC or epithelial SCC, they do not describe use of the

vaccine against other skin cancers, such as BCC or melanoma, likely based on the

understanding that BCC and melanoma are not associated with HPV infection.

[00013] New immunotherapy drugs have led to breakthroughs against many cancers but

there have been several failures over time. Among the newer drugs are CAR T-cells and

Immune Checkpoint Inhibitors (e.g., PD-1, PD-L1, and CTLA-4 inhibitors).

[00014] CAR T-cells have not performed well with solid tumors. Immune Checkpoint

Inhibitors (ICIs) have been used for solid tumors. One ICI, cemiplimab, has been approved

for advanced squamous cell carcinoma. Patients with complex skin squamous cell

carcinomas, such as dystrophic epidermolysis bullosa patients, appear not to respond as well

to cemiplimab which appears at least at some level due to immune based dysfunction.

[00015] Among the shortcomings of these ICI therapies are that they block limited (often

single) pathways. Combination of immunotherapies have produced better results, but the

benefits are limited due to their inhibition of a small number of specific pathways. With few

pathways blocked, treated cancers have also been shown to become resistant, no longer be

relying on these blocked pathways and finding other ways to survive.

[00016] There is a need in the medical and health fields for safe and efficacious cancer

treatments, including skin cancers or cancers that are typically not associated with HPV

infection, which are convenient for the patient as well as the health practitioner.

[00017] The limitations and disadvantages of the above-described treatments can be

overcome by the use of a composition or method in accordance with the subject invention.

[00018] An alternative approach to boosting these therapies, such as the use of one or

more ICI or other anti-cancer therapeutic in combination with a vaccine, which is inhibitory on

multiple levels, is a novel and needed advancement to the medical field.

PCT/US2021/054622

SUMMARY OF THE INVENTION

[00015] The subject invention concerns a method for treating a patient having skin

cancer or malignant tumor, whether or not associated with or related to human papilloma virus

(HPV) infection, said method comprising the steps of:

administering to a patient having cancer or in need of treatment of a tumor, skin

cancer or other cancerous lesion, a therapeutically effective dose of a commercially available

HPV vaccine. The vaccine can be administered directly to the cancer, tumor, or lesion, either

by direct application onto (topical) the tumor or lesion, or by direct injection into the tumor or

lesion. Alternatively, the vaccine can be administered for therapeutic use by systemic injection.

A method of treatment according to the subject invention can also include any combination of

topical application, direct or systemic injection. A therapeutically effective dose can be a

conventional, approved dose of the vaccine per its label indication.

[00016] In one embodiment, the method can comprise:

a) administering to a patient 27 years of age or older or a patient previously

not immunized with an HPV vaccine, a first dose of an HPV vaccine which is free of host-cell

peptide, polypeptide, or protein or a degradant product thereof;

b) administering to the patient a second dose of the HPV vaccine about one

month to about three months after the first administration; and

c) c) optionally, administering to the patient a third dose of the HPV vaccine

about five months to about seven months after the first dose.

[00020] Following the initial, conventional administration of the vaccine according to

step a), above, the second or third administrations according to steps b) and c), above, can be

by injection, or can be by topical administration of a composition comprising the vaccine.

Alternatively, the second or third administrations of steps b) or c) can include both injection

and by topical administration.

[00021] In one embodiment, the second dose of HPV vaccine is administered about two

months after administering the first dose and the third dose of HPV vaccine is administered

about six months after administering the first dose.

PCT/US2021/054622

[00022] The HPV vaccine can be selected from HPV quadrivalent (types 6, 11, 16, and

18) recombinant vaccine comprising HPV L1 proteins and HPV multivalent (types 16, 18, 31,

33, 45, 52, and 58) recombinant vaccine comprising HPV L1 proteins, and preferably is free

or substantially free of host-cell early antigen, e.g., E6 or E7.

[00023] In one preferred embodiment, the method does not comprise or is without

administering an additional or other immunostimulant or adjuvant.

[00024] In one preferred embodiment, the method comprises administering an additional

or other immunomodulatory agent, such as an immunostimulant or adjuvant. For purposes of

the subject invention, an immunomodulatory agent, such as an adjuvant, is generally not

regarded as a second active ingredient. Except where designated otherwise herein, an

immunomodulatory immunomodulatory agent agent or or adjuvant adjuvant is is distinguished distinguished from from aa second second active active ingredient ingredient used used in in

combination with HPV vaccine in accordance with the subject invention.

[00025] By carrying out the method, the size of the cancer or HPV-related lesion can be

substantially reduced, or completely eliminated. In addition, the incidence of recurrence of the

cancer or HPV-related lesion can be reduced. The method can be effective in treating or

reducing the incidence of recurrence of a cancer, benign tumor, or HPV-related lesion such as

squamous cell carcinoma, basal cell carcinoma, melanoma, verruca vulgaris, or condyloma

accuminata.

[00026] In one embodiment, the method can comprise a single dose of the vaccine. For

example, a single dose of the vaccine can be administered topically, or by injection directly

into a tumor or systemically to reduce the size or eliminate the tumor. A physician or healthcare

professional can administer a second or subsequent dose, as needed or as determined by the

physician or healthcare professional.

[00027] In one embodiment, the patient in need of treatment can be a person previously

immunized with the vaccine. In another embodiment, the patient in need of treatment can be a

person that has not been previously immunized with the vaccine.

[00028] Each dose of HPV vaccine administered in the above method steps is preferably

about 0.5 ml, and is more preferably 0.5 ml.

[00029] The method can further comprise establishing a positive diagnosis of cancer,

tumor, or HPV-related infection prior to administering the first dose of HPV vaccine.

[00030] An alternative embodiment of the method according to the subject invention

comprises treating a patient having cancer, tumor, or a human papilloma virus-related (HPV-

related) or HPV-unrelated cancerous lesion, wherein the method comprises administering a

dose of an HPV vaccine directly to the cancer, tumor, or lesion or an area immediately

surrounding the tumor or lesion.

[00031] This alternative embodiment of the method according to the subject invention

can further comprise the steps of:

administering a second dose of the HPV vaccine directly to the tumor or lesion or an

area immediately surrounding the tumor or lesion about one month to about three months after

administering the first dose; and

optionally, administering a third dose of the HPV vaccine directly to the tumor or lesion

or an area immediately surrounding the tumor or lesion about five months to about seven

months after administering the first dose.

[00032] These direct second or third administrations of a composition comprising the

vaccine can be topical applications, or can be by injection into the lesion.

[00033] In this alternative embodiment of the subject method, the second dose of HPV

vaccine can be administered about two months after administering the first dose and the third

dose of HPV vaccine can be administered about six months after administering the first dose.

[00034] By carrying out the alternative embodiment of the method according to the

subject invention, the size of the cancer, tumor, or HPV-related lesion can be substantially

reduced or completely eliminated. In addition, the incidence of recurrence of the cancer, tumor,

or HPV-related lesion can be reduced.

[00035] The preferred dose of each subsequent administration of HPV vaccine, if any,

is 0.5 ml.

[00036] The method according to any embodiment of the invention can be used for

treating cancer, tumor, or malignant HPV-related lesion, including, but not limited to, a tumor

associated or unassociated with HPV infection, squamous cell carcinoma, basal cell carcinoma,

and melanoma.

[00037] The method can further comprise establishing a positive diagnosis of cancer,

cancerous tumor, or HPV infection-associated or HPV-unrelated malignant lesion prior to

administering the first dose of HPV vaccine.

[00038] In one preferred embodiment, the direct or local administration of the vaccine

is administered by injection, and more preferably the method does not comprise administering

an additional or other immunostimulant or adjuvant, with, during or following the

administration of the vaccine.

[00039] Alternatively, the subject method can comprise administering an additional or

other immunomodulatory agent, e.g., and immunostimulant or adjuvant, with, during or

following the administration of the vaccine.

[00040] Yet another embodiment of the invention includes a composition comprising,

or a method of cancer treatment using, in combination, a first active agent which is an HPV

vaccine and a second active agent which can be an immunotherapy cancer drug, e.g., an

Immune Checkpoint Inhibitors, or "ICIs," such as PD-1, PD-L1, or CTLA-4 inhibitors, or

CAR T-cells. A second active ingredient in accordance with the subject invention can be a

semi-essential amino acid, such as N-acetyl cysteine (NAC.) Preferably, NAC is administered

orally, before, during or after injection of the vaccine composition.

[00041] Another second active useful in accordance with the subject invention can be a

messenger (m)RNA encoding a cytokine. Cytokine-encoding mRNA can produce one or more

interleukins, such as IL-12 or IL-15, interferon (e.g., IFN-a), granulocyte-macrophage colony- IFN-), granulocyte-macrophage colony-

stimulating factor, or the like. Effective antitumor activity of these cytokines involves multiple

immune cell populations and can be accompanied by intratumoral IFN-y induction, systemic IFN- induction, systemic

antigen-specific T cell expansion, increased granzyme B+ B TT cell cell infiltration, infiltration, and and formation formation of of

immune memory.

[00042] In one preferred embodiment, vaccine and mRNA encoding cytokine are

formulated for injection. In a more preferred embodiment for treating a cancerous tumor, the

WO wo 2022/081604 PCT/US2021/054622

vaccine and mRNA encoding cytokine can be formulated for intratumoral injection, and can

be injected sequentially, concomitantly, or in a fixed-dose combination formulation injected

together in a single composition.

[00043] In another preferred embodiment, the vaccine can be formulated for topical

administration and applied directly to the lesion in the form of a topical solution or suspension,

such as a liquid or spray, gel, cream, salve, ointment, foam or mousse, or the like.

[00044] The subject invention can particularly concern a method for treating a tumor

wherein the method comprises administering at least one dose of a commercially available

HPV vaccine to a patient having a tumor. Advantageously, the subject method has been found

to be effective for treating a tumor in glandular tissue, such as breast, pituitary (e.g., invasive

pituitary adenoma), prostate, or pancreas. This embodiment can include at administering at

least one dose of the vaccine directly into the tumor, itself.

[00045] The subject invention can comprise administering at least one dose of the

vaccine systemically, e.g., by intramuscular (IM) injection, alone, or in combination with

(concomitantly or shortly before or after) the direct administration of the vaccine to the tumor.

[00046] Alternatively, in certain instances, e.g., when the tumor presents on or near the

surface of the body, this method can further comprise topical administration of at least one dose

of the HPV vaccine, alone, or in combination with direct injection into the tumor or in

combination with systemic injection, or in combination with both direct and systemic injection.

[00047] Compositions comprising the vaccine are also included as part of the invention.

For example, the HPV vaccine can be used or formulated with one or more additional active

pharmaceutical ingredients for administration to the patient. Additional active pharmaceutical

ingredients can be one or more immunomodulatory agent for modulating the effect of the

vaccine, or one or more local anesthetic agent, e.g., lidocaine (with or without epinephrine),

for reducing patient discomfort during the injection. A preferred embodiment can include the

use of a first active agent which is an HPV vaccine and a second active agent which is an

immunotherapy cancer drug, e.g., an Immune Checkpoint Inhibitors such as PD-1, PD-L1, or

CTLA-4 inhibitors, or CAR T-cells. More preferably, the first and second active agents can be

formulated together in a fixed-dose combination composition, namely, a composition

comprising a therapeutically effective amount of an HPV vaccine such as GARDASIL,

PCT/US2021/054622

GARDASIL-9, CERVARIX, or the like, and a therapeutically effective amount of an Immune

Checkpoint Inhibitor or CAR T-cells. Most preferably, the first and second active agents are

formulated in an injectable dosage form.

[00048] One example of a composition of the invention comprises a 1:1 (v/v) ratio

mixture of 0.5 ml of a commercially available HPV vaccine and 0.5 ml of a commercially

available lidocaine solution (e.g., 0.5% (w/v), 1% (w/v), or 2% (w/v)). The composition can be

thoroughly mixed and injected into a patient for treatment. Ratios ranging from 1:10 (v/v)

vaccine:anesthetic solution vaccine:anesthetic solution to to 10:1 10:1 (v/v) (v/v) vaccine:anesthetic vaccine:anesthetic solution solution can can be be used, used, as as would would be be

understood in the art. When an mRNA encoding a cytokine is used in combination with the

vaccine, the mRNA encoding a cytokine can be admixed with the vaccine for injection or

admixed with the vaccine and additional active ingredient, such as a local anesthetic at standard

accepted doses.

[00049] The HPV vaccine can also be formulated with one or more excipients or diluents

for administration to the patient. Excipients and diluents can include one or more conventional

pharmaceutically acceptable ingredients useful for formulating topical preparations, including

but not limited to a bases for preparing a cream, emollient, gel, lotion, salve, or the like, and

can optionally include penetration enhancers, preservatives, release-controlling agents,

solubilizers, stabilizers, thickeners or thinners, or the like.

[00050] Solutions for injection can also include one or more buffer, emollient, diluent,

pH adjuster, preservative, solubilizer, stabilizer, or the like.

[00051] These compositions can be prepared as a manufactured product which can be

shipped, stored, and used as needed, including a later time, or can be compounded at the point

of care or remotely for immediate single-use treatment.

[00052] A composition of the invention can include one or more additional active

pharmaceutical ingredient without an excipient or diluent, or can include one or more active

pharmaceutical ingredient and one or more excipient or diluent.

[00053] A composition of the invention can include one or more excipient or diluent

without an additional active pharmaceutical ingredient, or can include one or more excipient

or diluent and one or more active pharmaceutical ingredient.

[00054]

[00054] To the To the knowledge knowledgeof of thethe inventor, inventor, administration administration of of HPVHPV vaccines vaccines comprising comprising 29 Jul 2025 2021360676 29 Jul 2025

only HPVantigens only HPV antigens(being (beingfree freeofofhost-cell host-cell peptides), peptides), to toaapreviously previouslyunimmunized patient, or unimmunized patient, or an an

adult patientaged adult patient aged2727 or or greater, greater, to to eliminate eliminate or reduce or reduce the incidence the incidence of recurrence of recurrence of skin cancer, of skin cancer,

tumor or other malignant skin lesion that is not an HPV-associated lesion, has not been previously tumor or other malignant skin lesion that is not an HPV-associated lesion, has not been previously

described. Nor has described. Nor hasthe thedirect direct or or local local administration administration of of aa vaccine vaccine bybytopical topicalapplication applicationoror by by direct direct injection injection into intothe thelesion lesionorortumor tumor been previously described been previously describedtoto eliminate eliminatethe thelesion lesion and and reduce the incidence of its recurrence. reduce the incidence of its recurrence. 2021360676

[00054a]

[00054a] Definitions Definitions of of specific specificembodiments ofthe embodiments of the invention invention as as claimed claimedherein hereinfollow. follow.

[00054b] According

[00054b] According to a embodiment, to a first first embodiment, there there is is provided provided a method a method of treating of treating a patient a patient

having skin having skin cancer, cancer, cancerous tumor,or cancerous tumor, or cancerous cancerouslesion, lesion, said said method comprisingadministering method comprising administering to the patient, a pharmaceutical composition consisting of: to the patient, a pharmaceutical composition consisting of:

aa human papillomavirus human papillomavirus (HPV) (HPV) vaccine vaccine selected selected fromfrom the group the group consisting consisting of anof an

HPVquadrivalent HPV quadrivalentrecombinant recombinant vaccine vaccine comprising comprising HPV HPV L1 proteins L1 proteins of HPVoftypes HPV6,types 11, 6, 1611, and16 and 18, 18, and and an an HPV multivalentrecombinant HPV multivalent recombinant vaccine vaccine comprising comprising HPV HPV L1 proteins L1 proteins of types of HPV HPV 16, types 16, 18, 31, 33, 18, 31, 33, 45, 45,52, 52,and and58;58; aa second active pharmaceutical second active ingredientwhich pharmaceutical ingredient whichisisananimmune-based immune-based therapeutic therapeutic

selected selected from an Immune from an Immune Checkpoint Checkpoint Inhibitor Inhibitor and and Chimeric Chimeric Antigen Antigen Receptor Receptor (CAR) T-cells; (CAR) T-cells;

and and

aa pharmaceutically acceptable pharmaceutically acceptable carrier. carrier.

[00054c] According

[00054c] According to second to a a second embodiment, embodiment, therethere is provided is provided an anticancer an anticancer pharmaceuticalcomposition pharmaceutical composition consistingof: consisting of: an HPVvaccine an HPV vaccineselected selected from from the the group group consisting consisting of of an an HPV quadrivalent HPV quadrivalent

recombinant vaccinecomprising recombinant vaccine comprising HPVHPV L1 protein L1 protein of types of HPV HPV 6,types 11, 6, 11,and 16, 16,18, andand18,anand HPV an HPV

multivalent recombinant multivalent recombinantvaccine vaccinecomprising comprising HPVHPV L1 protein L1 protein oftypes of HPV HPV 16, types 18,16, 31,18, 33,31, 45,33, 45, 52, 52, and and 58; 58;

aa second active pharmaceutical second active pharmaceuticalingredient ingredientwhich whichisisananimmune-based immune-based therapeutic therapeutic

selected from selected an Immune from an Immune Checkpoint Checkpoint Inhibitor Inhibitor andand CARCAR T-cells; T-cells; and and aa pharmaceutically acceptable pharmaceutically acceptable carrier. carrier.

[00054d]

[00054d] Any Any reference reference to publications to publications cited cited in thisinspecification this specification is an is not notadmission an admission that that the disclosures the disclosures constitute constitutecommon generalknowledge common general knowledgein in Australia Australia or or elsewhere. elsewhere.

11

[00054e]

[00054e] The term The term"comprise" “comprise”andand variantsof of variants thethe termterm suchsuch as “comprises” as "comprises" or or 29 Jul 2025 2021360676 29 Jul 2025

“comprising” "comprising" are are usedused herein herein to denote to denote the inclusion the inclusion of ainteger of a stated stated or integer statedor stated but integers integers not but not to exclude to anyother exclude any otherinteger integer or or any anyother otherintegers, integers, unless unless in in the the context of usage context of usage an anexclusive exclusive interpretation ofthe interpretation of theterm termis is required. required.

[Text

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11a 11a

DETAILED DESCRIPTION

[00055] The present invention is directed to a method of treating cancer, benign or

cancerous tumor, skin cancer, such as squamous cell carcinoma (SCC), or a cancerous skin

lesion associated with or unassociated with human papilloma virus (HPV) infection, and

includes treating a tumor originating in glandular tissue, such as breast, pituitary, prostate, or

pancreatic tissue. One embodiment of a method in accordance with the subject invention

comprises the administration of a commercially available HPV vaccine, such as an HPV

quadrivalent (types 6, 11, 16, and 18) recombinant vaccine, and a second active agent which is

an immunotherapy cancer drug, e.g., an Immune Checkpoint Inhibitors such as PD-1, PD-L1,

or CTLA-4 inhibitors, CAR T-cells, a semi-essential amino acid such as N-acetyl cysteine, or

an mRNA encoding a cytokine, to a patient having a cancer or tumor.

[00056] In one preferred embodiment, the subject method comprises administering at

least one dose of the HPV vaccine to a patent that has not been previously immunized with an

HPV vaccine, or to an adult patient aged 27 or older. For purposes of the subject invention, a

patient previously not immunized with an HPV vaccine is termed an "unimmunized patient"

regardless of other immunizations the patient may have received against other conditions or

diseases.

[00057] The dosing regimen can be a single administration by direct injection, systemic

injection, or topical application, or a combination of any of these administration routes.

Alternatively, the subject method can comprise multiple (more than one) administration, or

multiple (concomitant) administrations by direct injection, systemic injection, or topical

application of the vaccine.

[00058] The subject method can also comprise administering in accordance with the

conventionally accepted dosing series for a vaccine. For example, HPV vaccines are typically

administered using a dosing regimen comprising a first dose, a second dose about two months

following the first dose, and a third dose about six months following the first dose. These

second, third, or subsequent administrations can be systemic injection, e.g., conventional

intramuscular injection, or can be direct administration to the lesion by intralesional injection

or by topical administration.

[00059] The method embodiments of the present invention have surprisingly been found

to have beneficial results in treating, or minimizing the occurrence, recurrence, and/or progression of, cancer lesions or benign tumors that are not associated with HPV infection, such as basal-cell carcinoma (BBC) or melanoma.

[00060] While not being limited to any particular theory, it is proposed that the subject

method can increase, i.e., boost a patient's immune response that may manifest clinically as

increased surveillance in skin cells to decrease the likelihood of development and progression

of abnormal skin cells that produce the skin cancer, particularly, but not exclusively, SCC.

[00061] Alternatively, the method of the invention can interfere with inherent functional

activities of viral and virus-like proteins by other mechanisms. This interference would include

the complete or partial functional inactivation of viral and virus-like materials altered or

activated by exogenous and/or environmental agents such as ultraviolet light.

[00062] As used herein, the terms "HPV" and "human papillomavirus" refer to a non-

enveloped, double-stranded DNA viruses of the papillomavirus family. Their genomes are

circular and approximately 8 kilobase pairs in size. Most HPVs encode eight major proteins,

six located in the "early" region (E1-E2) and two in the "late" region (L1 (the major capsid

protein) and L2 (the minor capsid protein)). Over 120 HPV types have been identified, and

they are designated by numbers (e.g., HPV-16, HPV-18, etc.).

[00063] In one embodiment, an HPV vaccine of the subject invention comprises one or

more proteins (e.g., a recombinant L1 protein) from one, two, three, four, five, six, seven, eight,

nine, ten or more different HPV types. Methods of expressing HPV L1 proteins and methods

of making HPV vaccines are known in the art and described in, e.g., U.S. Patent Nos. 5,820,870

and 6,251,678, which are incorporated herein by reference in their entireties for all purposes.

[00064] In one embodiment, the HPV vaccine employed in the subject method contains

GARDASIL purified inactive viral or virus-like proteins, such as the commercially available GARDASIL®,

which is an HPV quadrivalent (types 6, 11, 16, and 18) recombinant vaccine or GARDASIL®

9, an HPV multivalent (types 16, 18, 31, 33, 45, 52, and 58) recombinant vaccine. In another

embodiment, the HPV vaccine is the commercially available CERVARIX®, which is an HPV

bivalent (types 16 and 18) recombinant vaccine. A vaccine useful in accordance with this

embodiment of the subject method is preferably free of host-cell and/or non-L1 HPV peptide,

polypeptide, or protein, such as the early antigens, E6 or E7, which are fragments of host-cell

peptides that present on the surface of an HPV-infected cell.

PCT/US2021/054622

[00065] The vaccine can be administered for treating cancerous or benign tumors,

including cancer lesions not associated with HPV infection, cancer (tumors or lesions)

associated with HPV infection, benign tumors not associated with HPV infection, or non-

cancerous HPV-related lesions in an unimmunized patient.

[00066] Alternatively, the vaccine can be administered to reduce the incidence ofof

recurrence of cancer, a benign tumor, or an HPV-related or HPV-unrelated lesion in an

unimmunized patient. In another embodiment, the vaccine can be administered to treat cancer,

benign tumor, or an HPV-related or HPV-unrelated lesion, or reduce the incidence of

recurrence thereof, in an adult patient aged 27 or greater.

[00067] More particularly, one preferred embodiment of the invention comprises a

method for the treatment of cancer, benign tumor, or HPV-related lesion, in a patient that is

unimmunized, or an adult patient aged 27 or older, comprising the steps of:

i. administering to the patient a first dose of an HPV recombinant vaccine free

of host-cell peptides, polypeptides, or proteins;

ii. administering to the patient a second dose of the HPV recombinant vaccine

free of host-cell peptides, polypeptides, or proteins between about one month and about three

months after the first dose; and

iii. iii. optionally, administering to the patient a third dose of the HPV vaccine free

of host-cell peptides, polypeptides, or proteins between about five months to about seven

months after administering the first dose.

[00068] The second or third, or subsequent, administration of the vaccine dose can be

systemic, e.g., intramuscular injection, or can be by direct administration to the lesion. The

direct administration of the vaccine composition to the lesion can be by intralesional injection,

or can be applied topically to the lesion. In a further embodiment, second, third or subsequent

administrations are both systemic and by direct application of vaccine to the lesion. Such direct

administration to the lesion can be intralesional injection or by topical application of a vaccine

composition formulated for topical administration.

[00069] It would be understood by medical practitioners that the reference to the timing

of subsequent administrations of the vaccine is approximate and can vary by days or even

weeks. This variation can result from patient compliance or non-compliance to the scheduled

dosing, clinical observation by the treating physician who may decide to advance (for more

WO wo 2022/081604 PCT/US2021/054622

aggressive treatment) or delay a subsequent administration for medical reasons. Generally,

however, an effective result can be achieved by following a dosing schedule where the second

dose is administered about two months following the first dose, and a third dose at about six

months after the first dose. Additional (fourth, or fifth) doses can be administered if the

physician deems that subsequent administrations can provide benefit to the patient.

[00070] A typical total dose for each administration according to the method of the

subject invention is about 0.5 ml of the vaccine, and is preferably 0.5 ml of a commercially

available HPV vaccine.

[00071] The terms "cancer," "cancerous," or "malignant" refer to or describe the

physiological condition in mammals that is typically characterized by unregulated cell growth.

Examples of cancer include but are not limited to: Cardiac: sarcoma (angiosarcoma,

fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma

and teratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiated small cell,

undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial

adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma; Gastrointestinal:

esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach

(carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma,

glucagonoma, gastrinoma, carcinoid tumors, vipoma), small bowel (adenocarcinoma,

lymphoma, carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma,

neurofibroma, fibroma), large bowel (adenocarcinoma, tubular adenoma, villous adenoma,

hamartoma, leiomyoma) colorectal; Genitourinary tract: kidney (adenocarcinoma, Wilm's

tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell

carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma),

testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma,

sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma);

Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma,

angiosarcoma, hepatocellular adenoma, hemangioma; Bone: osteogenic sarcoma

(osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's

sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant

cell tumor chordoma, osteochondroma (osteocartilaginous exostoses), benign chondroma,

chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors; Nervous

system: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges

PCT/US2021/054622

(meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma,

ependymoma, germinoma (pinealoma), glioblastoma multiform, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal cord neurofibroma, meningioma,

glioma, sarcoma); Gynecological: uterus (endometrial carcinoma), cervix (cervical carcinoma,

pre tumor cervical dysplasia), ovaries (ovarian carcinoma [serous cystadenocarcinoma,

mucinous cystadenocarcinoma, unclassified carcinoma], granulosa thecal cell tumors, Sertoli-

Leydig cell tumors, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma,

intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell

carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma),

fallopian tubes (carcinoma), breast; Hematologic: blood (myeloid leukemia (acute and

chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative

diseases, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's

lymphoma (malignant lymphoma); Skin: malignant melanoma, basal cell carcinoma,

squamous cell carcinoma, Kaposi's sarcoma, moles dysplastic nevi, lipoma, angioma,

dermatofibroma, keloids, psoriasis; and Adrenal glands: neuroblastoma. In another

embodiment, the cancer is carcinoma, lymphoma, leukemia, blastoma, and sarcoma. More

particular examples of such cancers include squamous cell carcinoma, myeloma, small-cell

lung cancer, non-small cell lung cancer, glioma, Hodgkin's lymphoma, non-Hodgkin's

lymphoma, acute myeloid leukemia (AML), multiple myeloma, gastrointestinal (tract) cancer,

renal cancer, ovarian cancer, liver cancer, lymphoblastic leukemia, lymphocytic leukemia,

colorectal cancer, endometrial cancer, kidney cancer, prostate cancer, thyroid cancer,

melanoma, chondrosarcoma, neuroblastoma, pancreatic cancer, glioblastoma multiforme,

cervical cancer, brain cancer, stomach cancer, bladder cancer, hepatoma, breast cancer, colon

carcinoma, and head and neck cancer. In certain exemplary embodiments, a cancer is an HPV-

associated cancer.

[00072] A particular example of cancer includes skin cancer, e.g., basal cell carcinoma

and/or squamous cell carcinoma, among other known skin cancers. Another example of cancer

includes breast cancer. Yet another example of cancer includes prostate cancer. Yet another

example includes penile cancer. Yet another example of cancer includes ovarian, cervical,

vaginal and/or vulvar cancer. Yet another example of cancer includes bladder cancer. Yet

another example of cancer includes colorectal and/or anal cancer. Yet another example of

cancer includes oropharyngeal cancer (e.g., cancer of the throat, soft palate, base of tongue,

PCT/US2021/054622

adenoids and/or tonsils). Yet another example of cancer includes renal cancer. Yet another

example of cancer includes liver cancer.

[00073] In certain exemplary embodiments, a cancer is associated with decreased

expression of Bcl-2-associated X protein (BAX) and/or Bcl-2 homologous antagonist/killer

(BAK1). In other exemplary embodiments, a cancer is associated with one or more aberrant

mitochondrial activities. In certain exemplary embodiments, an HPV vaccine of the invention

increases BAX and/or BAK1 expression in a tumor cell and/or promotes apoptosis of the tumor

cell. In other aspects, the combination of vitamin D and an HPV vaccine of the invention

increases BAX and/or BAK1 expression in a tumor cell and/or promotes apoptosis of a tumor

cell. In another embodiment, an HPV vaccine of the invention modulates one or more

mitochondrial activities in a tumor cell.

[00074] The above embodiments of a method of treatment according to the subject

invention can be efficacious for treating skin cancer in the patient, and particularly squamous

cell carcinoma, wherein a skin cancer lesion is reduced in size or eliminated following the three

administrations of the vaccine.

[00075] The treatment method in accordance with the subject invention can also reduce

the incidence of recurrence of benign tumors or cancer tumors or lesions, including skin cancer,

in the patient.

[00076] In particular the treatment method according to the subject invention comprises

eliminating, or reducing the size or incidence of recurrence of a cancerous tumor of the breast,

eliminating, or reducing the size or incidence of recurrence in a cancerous tumor of the prostate,

eliminating, or reducing the size or incidence of recurrence of a cancerous tumor of the

pancreas, or eliminating, or reducing the size or incidence of recurrence of a cancerous tumor

of the pituitary gland, e.g., invasive pituitary adenoma.

[00077] Other particular types of cancers or tumors that can benefit from treatment using

an HPV vaccine in accordance with the method of the subject invention include, and are not

limited to, cervical cancer, anal cancer, oropharyngeal cancers (throat, soft palate, base of

tongue, or tonsils), vaginal cancer, vulvar cancer, penile cancer, colorectal cancer, bladder

cancer, lung cancer, renal cancer, liver cancer, ovarian cancer, pancreatic mucinous cystic

neoplasms, gastric or stomach cancer.

WO wo 2022/081604 PCT/US2021/054622 PCT/US2021/054622

[00078] The method according to the subject invention can also be effective to reduce

the size or eliminate an HPV-associated, but non-cancerous, lesion, such as warts, including

genital warts, e.g., verruca vulgaris or condyloma accuminata

[00079] It is a further unexpected result of the present invention to provide a method of

reducing the incidence of recurrence of skin cancer, and particularly squamous cell carcinoma

following administration of one or more injections of HPV quadrivalent (types 6, 11, 16, and

18) recombinant vaccine, wherein the vaccine is substantially free of host-cell peptides,

polypeptides, or proteins which, as a result of HPV infection of the cell, present on the surface

of the infected cell. Further unexpected results of the subject method of treatment comprise

reducing the size of, eliminating, or reducing the incidence of recurrence of skin lesions that

are not associated with HPV infection, such as basal cell carcinoma or melanoma.

[00080] The invention pertains to uses of the above-described agents for the therapeutic

treatment of cancer. Accordingly, an HPV vaccine composition of the present invention is

incorporated into pharmaceutical compositions suitable for administration. Such compositions

typically comprise an HPV viral or viral-like protein and a pharmaceutically acceptable carrier.

As used herein the language "pharmaceutically acceptable carrier" is intended to include any

and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and

absorption delaying agents, and the like, compatible with pharmaceutical administration. The

use of such media and agents for pharmaceutically active substances is well known in the art.

Except insofar as any conventional media or agent is incompatible with the active compound,

use thereof in the compositions is contemplated. Supplementary active compounds can also be

incorporated into the compositions.

[00081] A pharmaceutical composition of the invention is formulated to be compatible

with its intended route of administration. Examples of routes of administration include

parenteral, e.g., intravenous (IV), intradermal, subcutaneous (SC or SQ), intraperitoneal,

intramuscular, oral (e.g., inhalation), transdermal (topical), and transmucosal administration.

Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can

include the following components: a sterile diluent such as water for injection, saline solution,

fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents;

antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic

acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such

as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium

WO wo 2022/081604 PCT/US2021/054622

chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or

sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable

syringes or multiple dose vials made of glass or plastic.

[00082] Pharmaceutical compositions suitable for injectable use include sterile aqueous

solutions (where water soluble) or dispersions and sterile powders for the extemporaneous

preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable

carriers include physiological saline, bacteriostatic water, Cremophor EL TM (BASF, (BASF,

Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be

sterile and should be fluid to the extent that easy syringe-ability exists. It must be stable under

the conditions of manufacture and storage and must be preserved against the contaminating

action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion

medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene

glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper

fluidity can be maintained, for example, by the use of a coating such as lecithin, by the

maintenance of the required particle size in the case of dispersion, or by the use of surfactants.

Prevention of the action of microorganisms can be achieved by various antibacterial and

antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and

the like. In many cases, it will be preferable to include isotonic agents, for example, sugars,

polyalcohols such as mannitol, sorbitol, sodium chloride in the composition. Prolonged

absorption of the injectable compositions can be brought about by including in the composition

an agent which delays absorption, for example, aluminum monostearate and gelatin.

[00083] Sterile injectable solutions can be prepared by incorporating the active

compound in the required amount in an appropriate solvent with one or a combination of

ingredients enumerated above, as required, followed by filtered sterilization. Generally,

dispersions are prepared by incorporating the active compound into a sterile vehicle which

contains a basic dispersion medium and the required other ingredients from those enumerated

above. In the case of sterile powders for the preparation of sterile injectable solutions, the

preferred methods of preparation are vacuum drying and freeze-drying which yields a powder

of the active ingredient plus any additional desired ingredient from a previously sterile-filtered

solution thereof.

[00084] Oral compositions generally include an inert diluent or an edible carrier. They

can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral

PCT/US2021/054622

therapeutic administration, the active compound can be incorporated with excipients and used

in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid

carrier to be swallowed or ingested as a solution or suspension, or for use as a mouthwash,

wherein the compound in the fluid carrier is applied orally and swished and expectorated or

swallowed. Pharmaceutically compatible binding agents, and/or adjuvant materials can be

included as part of the composition. The tablets, pills, capsules, troches and the like can contain

any of the following ingredients, or compounds of a similar nature: a binder such as

microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a

disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as

magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent

such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or

orange flavoring.

[00085] For administration by inhalation, the compounds are delivered in the form of an

aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g.,

a gas such as carbon dioxide, or a nebulizer.

[00086] Systemic administration can also be by transmucosal or transdermal means. For

transmucosal or transdermal administration, penetrants appropriate to the barrier to be

permeated are used in the formulation. Such penetrants are generally known in the art, and

include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid

derivatives. Transmucosal administration can be accomplished through the use of nasal sprays

or suppositories. For transdermal administration, the active compounds are formulated into

ointments, salves, gels, or creams as generally known in the art.

[00087] The compounds can also be prepared in the form of suppositories (e.g., with

conventional suppository bases such as cocoa butter and other glycerides) or retention enemas

for rectal delivery.

[00088] In one embodiment, the HPV viral or viral-like proteins are prepared with

carriers that will protect the compound against rapid elimination from the body, such as a

controlled release formulation, including implants and microencapsulated delivery systems.

Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate,

polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for

preparation of such formulations will be apparent to those skilled in the art. The materials can

WO wo 2022/081604 PCT/US2021/054622

also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc.

Liposomal suspensions (including liposomes targeted to infected cells with monoclonal

antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. These

can be prepared according to methods known to those skilled in the art, for example, as

described in U.S. Pat. No. 4,522,811.

[00089] It is especially advantageous to formulate oral or parenteral compositions in

dosage unit form for lease ofadministration ease of administrationand anduniformity uniformityof ofdosage. dosage.Dosage Dosageunit unitform formas asused used

herein refers to physically discrete units suited as unitary dosages for the subject to be treated;

each unit containing a predetermined quantity of active compound calculated to produce the

desired therapeutic effect in association with the required pharmaceutical carrier. The

specification for the dosage unit forms of the invention is dictated by and directly dependent

on the unique characteristics of the active compound and the particular therapeutic effect to be

achieved, and the limitations inherent in the art of compounding such an active compound for

the treatment of individuals.

[00090] Toxicity and therapeutic efficacy of such compounds can be determined by

standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for

determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose

therapeutically effective in 50% of the population). The dose ratio between toxic and

therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50/ED50.

Compounds that exhibit large therapeutic indices are preferred. Although compounds that

exhibit toxic side effects may be used, care should be taken to design a delivery system that

targets such compounds to the site of affected tissue in order to minimize potential damage to

uninfected cells and, thereby, reduce side effects.

[00091] The data obtained from the cell culture assays and animal studies can be used

in formulating a range of dosage for use in humans. The dosage of such compounds lies

preferably within a range of circulating concentrations that include the ED50 with little or no

toxicity. The dosage may vary within this range depending upon the dosage form employed

and the route of administration utilized. For any compound used in the method of the invention,

the therapeutically effective dose can be estimated initially from cell culture assays. A dose

may be formulated in animal models to achieve a circulating plasma concentration range that

includes the EC50 (i.e., the concentration of the test compound which achieves a half-maximal

response) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography.

[00092] The pharmaceutical compositions can be included in a container, pack, or

dispenser together with optional instructions for administration.

[00093] The route of delivery can be dependent on the disorder of the patient. In certain

exemplary embodiments, a subject diagnosed with skin cancer can be administered an HPV

vaccine composition of the invention by topical administration. In addition to an HPV vaccine

composition of the invention, a patient can be administered a second therapy, e.g., a palliative

therapy and/or disease-specific therapy. The secondary therapy can be, for example,

symptomatic (e.g., for alleviating symptoms), protective (e.g., for slowing or halting disease

progression), or restorative (e.g., for reversing the disease process). For the treatment of cancer,

for example, symptomatic therapies can further include another chemotherapeutic agent used

as a combination therapy as described further herein.

[00094] In general, an HPV vaccine composition of the invention can be administered

by any suitable method. As used herein, topical delivery can refer to the direct application of

an HPV vaccine composition to any surface of the body, including the eye, a mucous

membrane, surfaces of a body cavity, or to any internal surface. Formulations for topical

administration may include transdermal patches, ointments, lotions, creams, gels, drops,

sprays, and liquids. Conventional pharmaceutical carriers, aqueous, powder or oily bases,

thickeners and the like may be necessary or desirable. Topical administration can also be used

as a means to selectively deliver an HPV vaccine composition to the epidermis or dermis of a

subject, subject,orortoto specific strata specific thereof, strata or to an thereof, orunderlying tissue. to an underlying tissue.

[00095] Formulations for parenteral administration may include sterile aqueous

solutions which may also contain buffers, diluents, and other suitable additives. Intraventricular

injection may be facilitated by an intraventricular catheter, for example, attached to a reservoir.

For intravenous use, the total concentration of solutes should be controlled to render the

preparation isotonic.

[00096] An HPV vaccine composition of the invention can be administered to a subject

by pulmonary delivery. Pulmonary delivery compositions can be delivered by inhalation of a

dispersion SO so that the composition within the dispersion can reach the lung where it can be readily absorbed through the alveolar region directly into blood circulation. Pulmonary delivery can be effective both for systemic delivery and for localized delivery to treat diseases of the lungs.

[00097] Pulmonary delivery can be achieved by different approaches, including the use

of nebulized, aerosolized, micellular and dry powder-based formulations. Delivery can be

achieved with liquid nebulizers, aerosol-based inhalers, and dry powder dispersion devices.

Metered-dose devices are preferred. One of the benefits of using an atomizer or inhaler is that

the potential for contamination is minimized because the devices are self-contained. Dry

powder dispersion devices, for example, deliver drugs that may be readily formulated as dry

powders. An HPV vaccine composition may be stably stored as lyophilized or spray-dried

powders by itself or in combination with suitable powder carriers. The delivery of a

composition for inhalation can be mediated by a dosing timing element which can include a

timer, a dose counter, time measuring device, or a time indicator which when incorporated into

the device enables dose tracking, compliance monitoring, and/or dose triggering to a patient

during administration of the aerosol medicament.

[00098] The types of pharmaceutical excipients that are useful as carriers include

stabilizers such as Human Serum Albumin (HSA), bulking agents such as carbohydrates,

amino acids, and polypeptides; pH adjusters or buffers; salts such as sodium chloride; and the

like. These carriers may be in a crystalline or amorphous form or may be a mixture of the two.

[00099] Bulking agents that are particularly valuable include compatible carbohydrates,

polypeptides, amino acids, or combinations thereof. Suitable carbohydrates include

monosaccharides such as galactose, D-mannose, sorbose, and the like; disaccharides, such as

lactose, trehalose, and the like; cyclodextrins, such as 2-hydroxypropyl-B-cyclodextrin; and 2-hydroxypropyl--cyclodextrin; and

polysaccharides, such as raffinose, maltodextrins, dextrans, and the like; alditols, such as

mannitol, xylitol, and the like. A preferred group of carbohydrates includes lactose, trehalose,

raffinose maltodextrins, and mannitol. Suitable polypeptides include aspartame. Amino acids

include alanine and glycine, with glycine being preferred.

[000100] Suitable pH adjusters or buffers include organic salts prepared from organic

acids and bases, such as sodium citrate, sodium ascorbate, and the like; sodium citrate is

preferred.

[000101] One or more HPV viral or viral-like proteins of the invention (i.e., an HPV

vaccine) can be administered by oral or nasal delivery. For example, drugs administered

through these membranes have a rapid onset of action, provide therapeutic plasma levels, avoid

first pass effect of hepatic metabolism, and avoid exposure of the drug to the hostile

gastrointestinal (GI) environment. Additional advantages include easy access to the membrane

sites SO so that the drug can be applied, localized, and removed easily.

[000102] Another embodiment in accordance with the subject invention comprises

administering an HPV vaccine administered to a patient by direct or local administration, e.g.,

injection, into a skin lesion or surrounding area of the lesion. This direct administration method

can be useful in patients suffering from cancer, particularly skin cancer. This embodiment of

the method can also be useful for treating non-cancerous (benign) tumors, or non-cancerous

lesions associated with HPV, such as warts, e.g., verruca vulgaris or condyloma accuminata.

[000103] In an embodiment comprising direct injection into or surrounding a lesion, the

dosing regimen can comprise a single administration or more than one administration. For

example, a three-administration dosing series, as above, can be followed. Alternatively, a

physician can administer a subsequent dose as needed (prn) following an initial dose directly

into or surrounding the lesion. Divided dosing of the vaccine for any particular single time

point is considered to be a single administration.

[000104] This direct-administration embodiment of the invention can have beneficial

results in treating, or minimizing the occurrence, recurrence, and/or progression of, cancer

lesions or tumors such as basal-cell carcinoma (BBC) or melanoma, or non-cancerous (benign)

tumors that are not associated with HPV infection.

[000105] In one embodiment of the subject invention, the method is carried out without

the administration of an additional or other immunostimulant or adjuvant either with, during,

or following the treatment method of the invention.

[000106] Alternatively, the subject method can comprise administering an additional or

following the administration of the vaccine. Non-limiting examples of immunomodulatory

agents useful as part of the subject method include:

1) 1) Vitamin D and its analogues;

2) Sirolimus;

PCT/US2021/054622

3) Interferon and its analogues;

4) Vitamin A and its analogues, e.g., Soriatane (a retinoid)

5) Imiquimod; 6) Ingenol mebutate; and

7) T4 endonuclease 8) Antimetabolites, e.g., 5 Fluorouracil, Methotrexate

9) Cyclooxygenase inhibitors, e.g., Diclofenac

[000107] These agents can be given in combination locally or systemically with, or

contemporaneous with, the HPV vaccine as described herein, to enhance the effect of the

treatment. For example, in a previously HPV immunized patient having a tumor (skin, lung, or

the like), a combination of interferon and HPV antigen vaccine could be given locally.

Interferon may or may not also be given at the same time systemically. This administration can

enhance local destruction of the tumor or other lesion without the systemic side effects

associated with interferon.

[000108] In another aspect of the invention, the invention provides a method for treating

cancer in an individual comprising administering to the individual a combination therapy which

comprises an HPV vaccine and one or more additional chemotherapeutic agents other than the

HPV vaccine. The specific dosage and dosage schedule of the additional therapeutic agent can

further vary, and the optimal dose, dosing schedule and route of administration will be

determined based upon the specific therapeutic agent that is being used.

[000109] Examples of chemotherapeutic agents include alkylating agents such as thiotepa

and cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziri

dines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and

methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide,

triethylenethiophosphoramide and trimethylolomelamine; acetogenins (especially bullatacin

and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin;

cally statin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues);

cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin

(including the synthetic analogues, KW-2189 and CBI-TMI); eleutherobin; pancrati statin; a

sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine,

cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide

hydrochloride, hydrochloride, melphalan, melphalan, novembichin, novembichin, phenesterine, phenesterine, prednimustine, prednimustine, trofosfamide, trofosfamide, uracil uracil

mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as the enediyne antibiotics (e.g. calicheamicin, especially calicheamicin gammall and calicheamicin phill, see, e.g., Agnew, Chem. Intl. Ed. Engl., 33 : :

183-186 (1994); dynemicin, including dynemicin A; bisphosphonates, such as clodronate; an

esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne

antibiotic chromomophores), aclacinomysins, actinomycin, authramycin, azaserine,

bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including

morpholino- doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and cyanomorpholino-doxorubicin 2-pyrrolino-doxorubicin and

deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as

mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin,

puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex,

zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil (5-FU); folic

acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such

as fludarabine, 6- mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as

ancitabine, azacitidine, 6- azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine,

enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate,

epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane,

trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside;

aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine;

demecolcine; diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid; gallium

nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids such as maytansine and

ansamitocins; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet;

pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; razoxane;

rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2, 2',2"- trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and

anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol;

pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, e.g.

paclitaxel and doxetaxel; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine;

methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum;

etoposide (VP- 16); ifosfamide; mitoxantrone; vincristine; vinorelbine; novantrone; teniposide;

edatrexate; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor

RFS 2000; difluoromethylormthine (DMFO); retinoids such as retinoic acid; capecitabine; and

pharmaceutically acceptable salts, acids or derivatives of any of the above. Also included are

26

PCT/US2021/054622

anti-hormonal anti-hormonal agents agents that that act act to to regulate regulate or or inhibit inhibit hormone hormone action action on on tumors tumors such such as as anti- anti-

estrogens and selective estrogen receptor modulators (SERMs), including, for example,

tamoxifen, raloxifene, droloxifene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018,

onapristone, and toremifene (Fareston); aromatase inhibitors that inhibit the enzyme aromatase,

which regulates estrogen production in the adrenal glands, such as, for example, 4(5)-

imidazoles, aminoglutethimide, megestrol acetate, exemestane, formestane, fadrozole,

vorozole, letrozole, and anastrozole; and anti-androgens such as flutamide, nilutamide,

bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids or

derivatives of any of the above.

[000110] In one preferred embodiment, a method of treatment according to the invention

comprises administering a first active agent being an HPV vaccine as described herein, and a

second active agent which is an Immune Checkpoint Inhibitor or CAR T-cells. An Immune

Checkpoint Inhibitor can include a Programed Death 1 (PD-1) inhibitor; or a Programed Death

1 Ligand (PD-1L) inhibitor. Examples of PD-1 and PD-1L inhibitors are:

Cemiplimab Atezolizumab Atezolizumab Avelumab Bavencio Durvalumab Imfinzi

Keytruda Nivolumab Opdivo Pembrolizumab, and Tecentriq Tecentriq

[000111] An example of a Cytotoxic T-lymphocyte-Associated Protein 4 (CTLA-4)

inhibitor is Ipilumumab.

[000112] CAR T-cells is the abbreviation for Chimeric Antigen Receptor T cells and can

include a bispecific antibody, such as Catumaxomab, or a monoclonal antibody, such as

Campath, Brutuximab, Vismodigib, or the BRAF inhibitors, Vemurafenib, Dabrfenib, or

ecorafenib. In view of the limitations of these immune-based therapies, alone, or in

combination with one another, an HPV vaccine, being inhibitory on multiple levels, can

advantageously boost eh efficacy of the immune-based therapy. Vaccination to HPV infection

results in resistance to a naturally occurring, commonly encountered, infectious agent and can

PCT/US2021/054622

unexpectedly provide an immune response more complex than inhibition of a select few

pathways as produced by immune-based therapies, used alone. The response to vaccine seen

in elderly patients indicates an immune based effect in subjects with failing immune

surveillance function. Intralesional injection of HPV vaccine also indicates a direct local effect

on treated tumors.

[000113] Another second active ingredient useful for administration in combination with

HPV vaccine to a patient having cancer in accordance with the subject invention can be a semi-

essential amino acid, such as N-acetyl cysteine (NAC.) Preferably the NAC is administered

orally, before, during or after dosing of the vaccine composition via injection. NAC is typically

administered as an oral dosage form, 600-1200 mg per day, preferably 600 mg administered

orally, BID, and can be administered long-term, e.g., for the life of the patient, due to its low

toxicity.

[000114] In accordance with the subject invention, the NAC can be administered

following a complete series of HPV vaccine injections. Alternatively, NAC can be prescribed

for administration prior to the patient starting a complete series of HPV vaccine injections. The

time prior to HPV vaccine administration is not critical and can be 1 day up to one year prior

to the first HPV vaccine injection. Another alternative is to start the patient on NAC therapy

simultaneous with the initiation of the HPV vaccine injection series, for example, initiating

daily dosing of NAC on the first day of injecting a first HPV vaccine injection or at any time

during the course of the series of HPV vaccine injections.

[000115] In another embodiment of the subject invention, HPV vaccine can be administered in combination with administration of an mRNA encoding a cytokine. Cytokine-

encoding mRNA can produce one or more interleukins, such as IL-12 or IL-15, interferon (e.g.,

IFN-a), granulocyte-macrophage colony-stimulating IFN-), granulocyte-macrophage colony-stimulating factor, factor, or or the the like. like. Effective Effective antitumor antitumor

activity of these cytokines involves multiple immune cell populations and can be accompanied

by intratumoral IFN-y induction, systemic IFN- induction, systemic antigen-specific antigen-specific TT cell cell expansion, expansion, increased increased

granzyme B+ T cell infiltration, and formation of immune memory.

[000116] In one preferred embodiment, vaccine and mRNA encoding cytokine are

WO wo 2022/081604 PCT/US2021/054622

together in a single composition.

[000117] Each therapeutic agent in a combination therapy of the invention may be

administered either alone or in a medicament (also referred to herein as a pharmaceutical

composition or a fixed-dose combination product) which comprises the therapeutic agent and

one or more pharmaceutically acceptable carriers, excipients, and diluents, according to

standard pharmaceutical practice.

[000118] Each therapeutic agent in a combination therapy of the invention may be

administered simultaneously (i.e., in the same medicament), concurrently (i.e., in separate

medicaments administered one right after the other in any order) or sequentially in any order.

Sequential administration is particularly useful when the therapeutic agents in the combination

therapy are in different dosage forms (one agent is a tablet or capsule and another agent is a

sterile liquid) and/or are administered on different dosing schedules, e.g., a chemotherapeutic

that is administered at least daily and an HPV vaccine that is administered less frequently, such

as once weekly, once every two weeks, or once every three weeks.

[000119] In some embodiments, the HPV vaccine is administered before administration

of the chemotherapeutic agent, while in other embodiments, the HPV vaccine is administered

after administration of the chemotherapeutic agent. In another embodiment, the HPV vaccine

is administered concurrently with the chemotherapeutic agent.

[000120] In some embodiments, at least one of the therapeutic agents in the combination

therapy is administered using the same dosage regimen (dose, frequency, and duration of

treatment) that is typically employed when the agent is used as monotherapy for treating the

same cancer. In other embodiments, the patient receives a lower total amount of at least one of

the therapeutic agents in the combination therapy than when the agent is used as monotherapy,

e.g., smaller doses, less frequent doses, and/or shorter treatment duration.

[000121] Each therapeutic agent in a combination therapy of the invention can be

administered orally or parenterally, including the intravenous, intramuscular, intraperitoneal,

subcutaneous, rectal, topical, and transdermal routes of administration.

[000122] A combination therapy of the invention may be used prior to or following

surgery to remove a tumor and may be used prior to, during or after radiation therapy.

[000123] In some embodiments, a combination therapy of the invention is administered

to a patient who has not been previously treated with a biotherapeutic or chemotherapeutic

agent, i.e., is treatment-naive. In other embodiments, the combination therapy is administered

to a patient who failed to achieve a sustained response after prior therapy with a biotherapeutic

or chemotherapeutic agent, i.e., is treatment-experienced.

[000124] A combination therapy of the invention is typically used to treat a tumor that is

large enough to be found by palpation, visual observation or by imaging techniques well known

in the art, such as MRI, ultrasound, or CAT scan.

[000125] Any commercially available HPV vaccine can be employed for administration

directly to a cancer or HPV-related lesion. For example, this embodiment of the subject method

can comprise directly administering into or surrounding a lesion, a vaccine comprising purified

inactive viral or virus-like proteins, such as the commercially available GARDASIL®, which

is an HPV quadrivalent (types 6, 11, 16, and 18) recombinant vaccine or GARDASIL® 9, an GARDASIL 9, an

HPV multivalent (types 16, 18, 31, 33, 45, 52, and 58) recombinant vaccine or CERVARIX®,

an HPV bivalent (types 16 and 18) recombinant vaccine.

[000126] A vaccine useful in accordance with this embodiment of the subject method can

include host-cell peptides, polypeptides, or proteins, such as the early antigens, E6 or E7 or

exclude or be free of host-cell peptides, polypeptides, or proteins, such as the early antigens,

E6 or E7. The vaccine can be administered for treating cancer, a benign tumor, or HPV-related

lesion in a patient of any age, whether an unimmunized patient or a patient previously

immunized with an HPV vaccine.

[000127] The vaccine can be directly or locally administered into or surrounding a lesion

or tumor to reduce the incidence of recurrence of cancer, benign tumor, or an HPV-related

lesion in a patient.

[000128] In another embodiment, the vaccine can be administered to treat cancer, benign

tumor, or an HPV-related lesion, or reduce the incidence of recurrence thereof, in a patient up

to 26 years old (e.g., an infant, a child, an adolescent or a young adult) or, alternatively, an

adult patient aged 27 or greater.

[000129] More particularly, one preferred embodiment of the invention comprises a a

method for the treatment of cancerous or non-cancerous tumor or lesion in a patient comprising

WO wo 2022/081604 PCT/US2021/054622

the step of administering to the patient a dose of an HPV recombinant vaccine directly to the

lesion, tumor, or non-cancerous HPV-related lesion.

[000130] Alternatively, the method can comprise the following optional steps:

i. administering directly to a cancer lesion, benign tumor, or non-cancerous

HPV-related lesion of a patient a second dose of the HPV vaccine between about one month

and about three months after the first dose;

ii. administering directly to a cancer lesion, benign tumor, or non-cancerous

HPV-related lesion of a patient a subsequent dose of the HPV vaccine between about five

months to about seven months after administering the first dose; or

iii. administering directly to a cancer lesion, benign tumor, or non-cancerous

and about three months after the first dose, and administering directly to a cancer lesion, benign

tumor, or non-cancerous HPV-related lesion of a patient a subsequent dose of the HPV vaccine

between about five months to about seven months after administering the first dose.

[000131] It would be understood by medical practitioners that the reference to the timing

[000132] Selecting a dosage regimen (also referred to herein as an administration

regimen) depends on several factors, including the serum or tissue turnover rate of the entity,

the level of symptoms, the immunogenicity of the entity, and the accessibility of the target

cells, tissue or organ in the individual being treated. Preferably, a dosage regimen maximizes

the amount of therapeutic agent delivered to the patient consistent with an acceptable level of

side effects. Accordingly, the dose amount and dosing frequency depends in part on the

particular therapeutic agent, the severity of the cancer being treated, and patient characteristics.

Guidance in selecting appropriate doses of antibodies, cytokines, and small molecules is available. See, e.g., Wawrzynczak (1996) Antibody Therapy, Bios Scientific Pub. Ltd,

Oxfordshire, UK; Kresina (ed.) (1991) Monoclonal Antibodies, Cytokines and Arthritis,

Marcel Dekker, New York, NY; Bach (ed.) (1993) Monoclonal Antibodies and Peptide

Therapy in Autoimmune Diseases, Marcel Dekker, New York, NY; Baert et al. (2003) New

Engl. J. Med. 348:601-608; Milgrom et al. (1999) New Engl. J. Med. 341 : 1966-1973; Slamon

et al. (2001) New Engl. J. Med. 344:783-792; Beniaminovitz et al. (2000) New Engl. J. Med.

342:613-619; Ghosh et al. (2003) New Engl. J. Med. 348:24-32; Lipsky et al. (2000) New Engl.

J. Med. 343 : 1594-1602; Physicians' Desk Reference 2003 (Physicians' Desk Reference, 57th

Ed); Medical Economics Company; ISBN: 1563634457; 57th edition (November 2002).

Determination of the appropriate dosage regimen may be made by the clinician, e.g., using

parameters or factors known or suspected in the art to affect treatment or predicted to affect

treatment, and will depend, for example, the patient's clinical history (e.g., previous therapy),

the type and stage of the cancer to be treated and biomarkers of response to one or more of the

therapeutic agents in the combination therapy.

[000133] HPV viral or viral-like proteins of the invention may be administered by

continuous infusion, or by doses at intervals of, e.g., daily, every other day, three times per

week, or one time each week, two weeks, three weeks, monthly, bimonthly, etc. A total weekly

dose is generally at least 0.05 ug/kg, µg/kg, 0.2 ug/kg, µg/kg, 0.5 ug/kg, µg/kg, 1 ug/kg, µg/kg, 10 ug/kg, µg/kg, 100 ug/kg, µg/kg, 0.2

mg/kg, 1.0 mg/kg, 2.0 mg/kg, 10 mg/kg, 25 mg/kg, 50 mg/kg body weight or more. See, e.g.,

Yang et al. (2003) New Engl. J. Med. 349:427-434; Herold et al. (2002) New Engl. J. Med.

346: 1692-1698; Liu et al. (1999) J. Neurol. Neurosurg. Psych. 67:451-456; Portielji et al.

(20003) Cancer Immunol. Immunother. 52: 133-144.

[000134] In some embodiments, the dosing regimen will comprise administering the HPV

vaccine at a dose of 1, 2, 3, 5 or 10 mg/kg at intervals of about 14 days (+ (± 2 days) or about 21

(+ 2 days) or about 30 days (± days (± (+22days) days)or orabout aboutone oneweek week(+2 days), (± 2 two days), weeks two (+) weeks (±22days), days),

three weeks (+ (± 2 days) or four weeks (+ (± 2 days) throughout the course of treatment.

[000135] In other embodiments, the dosing regimen will comprise administering the HPV

vaccine at a dose of from about 0.005 mg/kg to about 10 mg/kg, with intra-patient dose

escalation. In other escalating dose embodiments, the interval between doses will be

progressively shortened, e.g., about 30 days (+2 (±2 days) between the first and second dose, about

14 days (+ (± 2 days) between the second and third doses. In certain embodiments, the dosing

interval will be about 14 days (+ (± 2 days), for doses subsequent to the second dose. A typical

WO wo 2022/081604 PCT/US2021/054622

total dose for each direct or local administration according to the method of the subject

invention is about 0.5 ml of the vaccine, e.g., of a commercially available vaccine. Each 0.5 ml

dose can be administered, e.g., by intralesional injection, as a bolus of the entire 0.5 ml or can

be administered as a divided dose as a plurality of 0.1-0.2 ml partial administrations into the

lesion, an area surrounding the lesion, or both.

[000136] According to certain embodiments, multiple doses of an HPV vaccine may be

administered to a subject over a defined time course. The methods include, for example,

sequentially administering to a subject multiple doses of an HPV vaccine. As used herein,

"sequentially administering" means that each dose of an HPV vaccine is administered to the

subject at a different point in time, e.g., on different days separated by a predetermined interval

(e.g., hours, days, weeks, or months). The present invention includes methods which comprise

sequentially administering to the patient a single initial dose of an HPV vaccine, followed by

one or more secondary doses of an HPV vaccine, and optionally followed by one or more

tertiary doses of an HPV vaccine.

[000137] The terms "initial dose," "secondary doses," and "tertiary doses," refer to the

temporal sequence of administration of an HPV vaccine. Thus, the "initial dose" is the dose

which is administered at the beginning of the treatment regimen (also referred to as the

"baseline dose"); the "secondary doses" are the doses which are administered after the initial

dose; and the "tertiary doses" are the doses which are administered after the secondary doses.

The initial, secondary, and tertiary doses may all contain the same amount of an HPV vaccine

(e.g., of the one or more HPV viral or viral-like proteins), but will generally differ from one

another in terms of frequency of administration. In certain embodiments, however, the amount

of an HPV vaccine (e.g., of the one or more HPV viral or viral-like proteins) contained in the

initial, secondary and/or tertiary doses will vary from one another (e.g., adjusted up or down

as appropriate) during the course of treatment.

[000138] In one exemplary embodiment, each secondary and/or tertiary dose is

administered administered 1 to 14 (e.g., 1 to 1, 11/2, 14 (e.g., 2, 21/2, 1, 1½, 3, 31/2, 2, 2½, 3, 3½,4, 4, 41/2, 4½,5,5,51/2, 5½, 6, 6, 61/2, 7, 71/2, 6½, 7, 7½, 8,8, 8½, 81/2, 9,9,9½, 91/2, 10,10,

101/2, 11, 11½, 10½, 11, 11 1/2, 12,12, 12 1/2, 12½, 13, 131/2, 13, 13½, 14, 141/2, 14, 14½, or more) or more) weeksweeks after after thethe immediately preceding immediately preceding

dose. In another exemplary embodiment, each secondary and/or tertiary dose is administered

1 1 toto14 14 (e.g., 1, 11/2, (e.g., 1,2,1½, 21/2, 2,3, 2½, 31/2,3, 4, 3½, 41/2, 4, 5, 51/2, 4½, 6, 5,61/2, 5½, 7,6,7 1/2, 6½, 8,7,81/2, 7½,9,8, 91/2, 8½,10,9, 101/2, 9½,11, 10,111/2, 10½, 11, 11½,

12, 12 1/2, 12½, 13,13, 131/2, 13½, 14, 141/2, 14, 14½, or months or more) more) months after after the the immediately immediately preceding preceding dose. dose. The The phrase phrase

"the immediately preceding dose," as used herein, means, in a sequence of multiple

WO wo 2022/081604 PCT/US2021/054622

administrations, the dose of an HPV vaccine which is administered to a patient prior to the

administration of the very next dose in the sequence with no intervening doses.

[000139] These methods may include administering to a patient any number of secondary

and/or tertiary doses of an HPV vaccine. For example, in certain embodiments, only a single

secondary dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4,

5, 6, 7, 8, or more) secondary doses are administered to the patient. Likewise, in certain

embodiments, only a single tertiary dose is administered to the patient. In other embodiments,

two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) tertiary doses are administered to the patient.

[000140] In embodiments involving multiple secondary doses, each secondary dose may

be administered at the same frequency as the other secondary doses. For example, each

secondary dose may be administered to the patient 1 to 3 months after the immediately

preceding dose. Similarly, in embodiments involving multiple tertiary doses, each tertiary dose

may be administered at the same frequency as the other tertiary doses. For example, each

tertiary dose may be administered to the patient 1 to 3 months after the immediately preceding

dose. Alternatively, the frequency at which the secondary and/or tertiary doses are administered

to a patient can vary over the course of the treatment regimen. The frequency of administration

may also be adjusted during the course of treatment by a physician depending on the needs of

the individual patient following clinical examination.

[000141] In certain embodiments, the initial dose (e.g., a "loading dose") is higher than

either or both of the secondary and tertiary doses. For example, the initial dose can be a loading

dose, which is 1.5x, 2x, 2.5x, 3x or more, greater than the secondary dose.

[000142] The above direct or local administration method of treatment can be efficacious

for treating skin cancer in the patient, and particularly squamous cell carcinoma, wherein a skin

cancer lesion is reduced in size or eliminated following the three administrations of the vaccine.

[000143] The direct or local administration treatment method according to the subject

invention can also reduce the incidence of recurrence of cancer, including skin cancer, in the

patient.

[000144] The direct or local administration method can also be effective to reduce the

size or eliminate a benign tumor, whether or not associated with HPV infection, or an HPV-

PCT/US2021/054622

associated, but non-cancerous, lesion, such as warts, including genital warts, e.g., verruca

vulgaris or condyloma accuminata.

[000145] The direct or local administration method can also be effective to reduce the

incidence of recurrence of a benign tumor, whether or not associated with HPV infection, or

an HPV-associated, but non-cancerous, lesion, such as warts, including genital warts, e.g.,

verruca vulgaris or condyloma accuminata.

[000146] It is a further unexpected result of the present invention to provide a method of

eliminating or reducing the size or incidence of recurrence of skin cancer, and particularly

squamous cell carcinoma following direct or local administration of one or more injections of

HPV bivalent (types 16 and 18) recombinant vaccine, HPV quadrivalent (types 6, 11, 16, and

18) recombinant vaccine or an HPV multivalent (types 16, 18, 31, 33, 45, 52, and 58)

recombinant recombinantvaccine. vaccine.

[000147] Further unexpected results of the subject direct or local administration method

of treatment comprise reducing the size of, eliminating, or reducing the incidence of recurrence

of skin lesions that are not associated with HPV infection, such as basal cell carcinoma or

melanoma.

[000148] In one embodiment of the subject invention, the direct or local administration

method is carried out without the administration of an additional or other immunostimulant or

adjuvant.

[000149] In certain embodiments, the subject method can comprise administering an

additional or other immunomodulatory agent, e.g., and immunostimulant or adjuvant, with,

during or following the administration of the vaccine. Non-limiting examples of

immunomodulatory agents useful as part of the subject method include:

1) Vitamin D and its analogues;

2) Sirolimus;

3) Interferon and its analogues;

4) Vitamin A and its analogues, e.g., Soriatane (a retinoid)

5) Imiquimod; 6) Ingenol mebutate; and

7) T4 endonuclease 8) Antimetabolites, e.g., 5 Fluorouracil, Methotrexate

9) cyclooxygenase inhibitors, e.g., Diclofenac

[000150]

[000150] TheseThese agents agents cangiven can be be given in combination in combination locally locally or systemically or systemically with, with, or or 29 Jul 2025 2021360676 29 Jul 2025

contemporaneous with, contemporaneous with, thethe HPV HPV vaccine vaccine as described as described herein, herein, to enhance to enhance theofeffect the effect the of the treatment. For treatment. example,inin aa previously For example, previouslyHPV HPV immunized immunized patient patient having having a tumor a tumor (skin, (skin, lung,lung, or or the like), a combination of interferon and HPV antigen vaccine could be given locally. Interferon the like), a combination of interferon and HPV antigen vaccine could be given locally. Interferon

mayorormay may may not not alsobebe also given given at at thethe same same time time systemically. systemically. ThisThis administration administration can can enhance enhance

local local destruction destruction of of the the tumor or other tumor or other lesion lesion without the systemic without the side effects systemic side effects associated associated with with

interferon. interferon. 2021360676

[000151]

[000151] Topical Topical application application canbeneficial can be be beneficial for for several several reasons, reasons, including including the the elimination of infection elimination of infection risk risk caused caused by injection, but by injection, but can can also also be be advantageous bywide-spread advantageous by wide-spread application overlarge application over large areas areas in in order order to treat to treat precancerous precancerous (actinic (actinic keratoses) keratoses) as wellas aswell as malignant malignant

tumors. In tumors. In addition, addition, the thetopical topicaladministration administrationcan canprovide providecosmetic cosmetic enhancement ofthe enhancement of the skin, skin, by by

decreasing that appearance of pigment irregularities, poikiloderma, and scaling. decreasing that appearance of pigment irregularities, poikiloderma, and scaling.

[000152] Embodiments

[000152] Embodiments of the invention of the subject subject invention advantageously advantageously provide a cost-effective, provide a cost-effective,

safe, safe, efficacious, efficacious,and andconvenient convenient treatment treatment for for reducing or ameliorating reducing or the growth ameliorating the growthororsize size of of aa cancer tumororor lesion, cancer tumor lesion, including including a a skin skin cancer cancer lesion lesion such such as as SCC, BCCorormelanoma SCC, BCC melanoma tumor tumor or or

lesion. Embodiments lesion. Embodiments of of thethe subject subject invention invention also also advantageously advantageously provide provide a cost-effective, a cost-effective,

efficacious, efficacious, and convenienttreatment and convenient treatment forfor curing curing skinskin cancer cancer lesions, lesions, and aand a cost-effective, cost-effective,

efficacious, efficacious, and and convenient methodtotoreduce convenient method reduce thethe incidence incidence of of recurrence recurrence of of cancer, cancer, including including

skin cancerlesions. skin cancer lesions.

[000153]

[000153] The The subject subject method method of treating of treating or reducing or reducing the incidence the incidence of recurrence of recurrence of skin of skin cancer comprisesadministering cancer comprises administering an an HPV HPV vaccine vaccine in onein orone moreordoses moretodoses to a patient. a patient. In one In one

embodiment, themethod embodiment, the method includes includes administration administration of aoffirst a firstdose dose of of HPVHPV quadrivalent quadrivalent (types (types 6, 6,

11, 11, 16, 16, and and 18) recombinantvaccine 18) recombinant vaccinetotoa apatient, patient, aa second seconddose doseofofHPV HPV quadrivalent quadrivalent (types (types 6, 6,

11, 11, 16, 16, and 18) recombinant and 18) recombinantvaccine vaccineapproximately approximately two two months months thereafter, thereafter, and and a third a third dosedose of of

HPVquadrivalent HPV quadrivalent(types (types6,6,11, 11, 16, 16, and 18) recombinant and 18) vaccineapproximately recombinant vaccine approximately four four months months after after

the second the dose. In second dose. In aa preferred preferred embodiment, eachdose embodiment, each doseisis0.5 0.5 ml. ml.

[000154]

[000154] Thesubject The subjectmethod methodcancan be advantageous be advantageous in it in that thatcanit be canperformed be performed using ausing a commercially availableHPV commercially available HPV bivalent bivalent (types (types 1616 and and 18)18) recombinant recombinant vaccine, vaccine, HPV HPV quadrivalent quadrivalent

(types 6, 11, (types 6, 11, 16, and 18) 16, and 18) vaccine vaccineororHPV HPV multivalent multivalent (types (types 16, 16, 18, 18, 31, 31, 33, 52, 33, 45, 45, and 52, 58) and 58) recombinantvaccine recombinant vaccineasasa atherapeutic therapeuticagent agentrather ratherthan thanororininaddition additiontoto its its use use as as a a preventive preventive

vaccine. vaccine.

36

[000155] A preventive vaccine is understood to be a vaccine composition administered

prior to exposure to or infection with an agent such as human papilloma virus (HPV).

Preventive vaccines for protection against or prevention of HPV infection and associated

cancers are commercially available are therefore known to be safe. GARDASIL® is an GARDASIL is an HPV HPV

quadrivalent (types 6, 11, 16, and 18) recombinant vaccine and GARDASIL® 9, is GARDASIL 9, is an an HPV HPV

multivalent (types 16, 18, 31, 33, 45, 52, and 58) recombinant vaccine currently marketed as a

preventive vaccine in the United States by Merck & Co., Inc. Whitehouse Station, NJ 08889

USA. CERVARIX® is an HPV bivalent (types 16 and 18) recombinant vaccine available from

GlaxoSmithKline (Brentford, England).

[000156] By use of a commercially available vaccine, the vaccine can be readily accessed

by a physician or healthcare practitioner. Moreover, the use of an HPV bivalent (types 16 and

18) recombinant vaccine, an HPV quadrivalent (types 6, 11, 16, and 18) recombinant vaccine

or HPV multivalent (types 16, 18, 31, 33, 45, 52, and 58) recombinant vaccine in accordance

with the subject method do not require secondary or additional immunostimulants or adjuvants.

These commercially available HPV bivalent (types 16 and 18), HPV quadrivalent (types 6, 11,

16, and 18) or HPV multivalent (types 16, 18, 31, 33, 45, 52, and 58) recombinant vaccines are

free, or substantially free, of host-cell and/or non-L1 viral peptides, polypeptides, or proteins,

such as the antigens, E6 or E7.

[000157] Advantageously, the unexpected result of treating cancer, benign tumor, or

HPV-related skin lesions, including skin cancers that are associated with HPV infection or skin

cancers that are not associated with HPV infection, can be achieved using the subject method

as described herein.

[000158] Another embodiment of the subject invention includes a composition for

carrying out a method of treatment as described. Compositions comprising the vaccine and an

added ingredient - one or more of an active pharmaceutical ingredients, excipient, or diluents,

for example - are also included as part of the invention. In a composition of the invention, HPV

vaccine can be formulated with one or more additional active pharmaceutical ingredients for

administration to the patient. Additional active pharmaceutical ingredients can be one or more

immunomodulatory agent for modulating the effect of the vaccine, or one or more local

anesthetic agent, e.g., lidocaine (with or without epinephrine), for reducing patient discomfort

during the injection.

PCT/US2021/054622

[000159] One embodiment of a composition of the subject invention comprises

commercially available HPV vaccine formulated with one or more immunomodulatory agent.

The one or more immunomodulatory agent can be selected from the group consisting of:

1) Vitamin D and its analogues;

2) Sirolimus;

3) Interferon and its analogues;

4) Vitamin A and its analogues, e.g., Soriatane (a retinoid)

5) Imiquimod; 6) Ingenol mebutate; Ingenol mebutate;andand

7) T4 endonuclease 8) Antimetabolites, e.g., 5 Fluorouracil, Methotrexate

9) cyclooxygenase inhibitors, e.g., Diclofenac

[000160] A composition comprising HPV vaccine and at least one immunomodulatory

agent can advantageously provide enhanced effect of the anti-cancer therapeutic activity of the

HPV vaccine.

[000161] In another embodiment, a composition according to the invention comprises

HPV vaccine formulated with an mRNA encoding a cytokine in a single, fixed dose

combination composition.

[000162] One embodiment of a composition of the subject invention comprises commercially available HPV vaccine formulated with one or more local anesthetic agent. The

one or more local anesthetic agent can be selected from the group consisting of: the ester local

anesthetics, namely procaine, benzocaine, chloroprocaine, cocaine, cyclomethycaine,

dimethocaine/larocaine, piperocaine, propoxycaine, procaine, proparacaine, and tetracaine, or

the amide local anesthetics, namely, lidocaine, articaine, bupivacaine, cinchocaine, etidocaine,

levobupivacaine, lignocaine, mepivacaine, prilocaine, ropivacaine, and trimecaine.

[000163] One example of a composition of the invention comprises a 1:1 (v/v) ratio

vaccine:anesthetic solution to 10:1 (v/v) vaccine:anesthetic solution can be used, as would be

understood in the art.

WO wo 2022/081604 PCT/US2021/054622

[000164] One embodiment of a composition of the subject invention comprises commercially available HPV vaccine formulated in a fixed-dose combination product with one

or more anti-cancer agent selected from the group consisting of an Immune Checkpoint

Inhibitors and CAR T-cells.

[000165] Immune Checkpoint Inhibitors of the invention include, but are not limited to,

the PD-1/PD-L1 Inhibitors Cemiplimab, Atezolizumab, Avelumab, Bavencio, Durvalumab,

Imfinzi, Keytruda, Nivolumab, Opdivo, Pembrolizumab, and Tecentriq, or can include the

CRLA-4 inhibitor Ipilumumab. CAR T-cells that can be formulated in a fixed-dose

combination product with HPV vaccine include bispecific antibodies, such as Catumaxomab;

monoclonal antibodies, such as Campath, Brutuximab, or Vismodigib, or the BRAF inhibitors,

Vemurafenib, Dabrfenib, or ecorafenib.

[000166] The HPV vaccine and second active agent such as Immune Checkpoint Inhibitors and CAR T-cells can also be formulated with one or more excipients or diluents for

administration to the patient. Solutions for injection can also include one or more buffer,

emollient, diluent, emollient, pH adjuster, diluent, preservative, pH adjuster, solubilizer, preservative, stabilizer, solubilizer, or the like.or the like. stabilizer,

[000167] Excipients and diluents can include one or more conventional pharmaceutically

acceptable ingredients useful for formulating topical preparations, including but not limited to,

a base for preparing a cream, emollient, gel, lotion, salve, or the like, and can optionally include

penetration enhancers, preservatives, release-controlling agents, solubilizers, stabilizers,

thickeners or thinners, or the like.

[000168] A topical composition comprising a vaccine useful in accordance with the

subject invention can be formulated as is conventionally known in the pharmaceutical arts, and

can comprise one or more additional ingredients or excipients, such as an organic or inorganic

solvent (aqueous or non-aqueous), stabilizing agent, penetration enhancer, buffer, gelling

agent, polymeric agent, lubricant, glidant, cream, wax, suspending agent, surfactant, or the like.

The formulation can further include a penetration enhancer, such as DMSO. The formulation

can be provided as a topical solution, lotion or shake lotion, ointment, cream, gel, foam,

transdermal patch, biofrequency chip, powder, solid, sponge, tape, paste, tincture, micelle or

liposome, or the like.

PCT/US2021/054622

[000169] These compositions can be prepared as a manufactured product which can be

of care or remotely for immediate single-use treatment.

[000170] A composition of the invention can include one or more additional active

pharmaceutical ingredient and one or more excipient or diluent.

[000171] A composition of the invention can include one or more excipient or diluent

or diluent and one or more active pharmaceutical ingredient

EXAMPLES Example 1 - skin cancer

[000172] The following charts provide the results from the subject method of treatment

carried out in three patients experiencing relatively frequent recurrence rates of skin cancer,

including squamous cell carcinoma (SCC) as well as basal-cell carcinoma.

[000173] The data presented below represents an average number of distinctive

recurrences of skin cancer per month for a period of time prior to and after undergoing the

method of treatment described herein.

A. Patient 1

[000174] Patient 1 was administered three 0.5ml doses, including a first 0.5ml dose, a

second 0.5ml dose two months later, and a third 0.5ml dose four months after the second dose.

In a follow-up exam three months after administration of the third dose of HPV quadrivalent

(types 6, 11, 16, and 18) recombinant vaccine, Patient 1 had experienced zero recurrences of

skin cancer, including both SCC and BCC types, during the three-month period. Prior to

commencement of the treatment method, Patient 1 had more than 300 distinctive occurrences

of skin cancer during his lifetime.

PATIENT 1 Time Period SCC BCC (Months) Prior to Commencement 16 1.80 0.25 of Treatment Method After Commencement 16 0.37 0.00 of Treatment Method

WO wo 2022/081604 PCT/US2021/054622 PCT/US2021/054622

B. Patient 2

[000175] Patient 2 was administered three 0.5ml doses of HPV quadrivalent (types 6, 11,

16, and 18) recombinant vaccine, including a first 0.5 ml dose, a second 0.5ml dose two

months later, and a third 0.5ml dose four months after the second dose.

PATIENT 2 Time Period SCC SCC BCC (Months) Prior to Commencement 13 2.07 0.53 of Treatment Method After After Commencement Commencementof of 13 0.23 0.3

Treatment Method

C. Patient 3

[000176] Patient 3 was administered three 0.5ml doses of HPV quadrivalent (types 6, 11,

16, and 18) recombinant vaccine, including a first 0.5ml dose, a second 0.5ml dose two

months later, and a third 0.5ml dose eight months after the second dose.

PATIENT 3 Time Period SCC BCC (Months) Prior to Commencement 22 0.18 0.13 of Treatment Method After After Commencement Commencementof of 22 0.09 0.04 Treatment Method

[000177] As a group, each of the patients who underwent the method of treatment using

HPV quadrivalent (types 6, 11, 16, and 18) recombinant vaccine experienced a significant

decrease in the number of skin cancer recurrences, as well as improvement in the texture and

appearance of the skin with decreased scaling and an increase in general skin suppleness.

[000178] Generally, the method of treatment described herein serves to effectively

increase, i.e., boost, the patient's immune surveillance in skin cells in order to decrease the

likelihood of a development of abnormal skin cells that produce the skin cancer. The method

of the present invention has been shown to treat and prevent recurrence of SCC, and to

significantly reduce recurrence of BCC. It is also possible that the increase in immune

surveillance, as a result of the treatment method, will concomitantly decrease the incidence of

malignant melanoma.

WO wo 2022/081604 PCT/US2021/054622

[000179] In one embodiment, the method of treatment for eliminating or reducing the

incidence of recurrence of skin cancer includes administering the HPV quadrivalent (types 6,

11, 16, and 18) recombinant vaccine in the form of an injection directly into the cancerous

tissue or an area of tissue immediately surrounding the cancerous tissue.

Example 2 - Breast cancer

[000180] A previously HPV-vaccinated 32-year old woman with no history of breast

cancer, no family history, and no risk factors, was diagnosed with metastatic breast cancer. Her

main tumor was measured by ultrasound as being about 4.1 centimeters in diameter. These

metastatic tumors can double in size in about 12 weeks.

[000181] With the patient's fully informed consent and knowledge, the tumor was

directly injected with a standard initial dose (about 0.5 ml) of a commercially available HPV

vaccine. A second dose 0.5 ml, diluted with saline and lidocaine to about 3 ml, was directly

administered to the tumor about two weeks after the first injection. At that time, it was harder

to find the tumor to inject, and was believed to have been reduced in size.

[000182] A follow up ultrasound recently showed that the tumor had been reduced in size

to about 2.7 centimeters in diameter, corresponding to an approximate 35% reduction in

diameter, and a 75% reduction in tumor volume (volume for a sphere is calculated as 4/3 pi X x

radius cubed).

[000183] With the expected doubling of size, the tumor should have increased by about

40%. 40%. to to aa tumor tumor diameter diameter of of about about 4.6 4.6 centimeters. centimeters.

Example 3 - metastatic basosquamous carcinoma

[000184] A 99-year old female presented with metastatic basosquamous carcinoma on

the leg. metastatic basosquamous carcinoma, severe enough that she was referred to

dermatology for palliative treatment and no further options were available but amputation of

the limb to prevent further spreading of the cancer.

[000185] A single injection of a conventional dose (about 0.5 ml) of a commercially

available HPV vaccine was administered to the patient, intramuscularly (systemically).

Additional standard doses of the HPV vaccine were injected into each of two or more sites of

the larger lesions.

WO wo 2022/081604 PCT/US2021/054622

[000186] Within four weeks of the treatment with HPV vaccine, the lesions were

substantially visually improved, and the cancer had no further spreading on the leg. The patient

is currently in remission from further or increased size of the lesions.

Example 4 - penile cancer

[000187] A 45 year old HIV-positive man with a two-year history of squamous cell

carcinoma of the penis that was recalcitrant to treatment with a variety of topical and surgical

methods methodswas wastreated with treated three with equalequal three doses doses of GARDASIL®, , intramuscularly, of GARDASIL, in accordance intramuscularly, in accordance

with the label instructions.

[000188] Within four days, the patient's pain started to lessen, from a pain scale rating of

9-10 on a 10-scale rating to zero over the course of several weeks

[000189] Recent examinations with confocal microscopy show no evidence of malignancy. Confocal photography can be used to detect cancer on skin without the need for

biopsy.

Example 5 - aggressive squamous cell carcinoma

[000190] An aggressive rapidly growing recurrent squamous cell carcinoma on the lower

extremity of an elderly man with history of renal cell carcinoma, and history of chemotherapy

was treated with two intralesional injections of GARDASIL® mixed with lidocaine 1% with

epinephrine.

[000191] The patient had previously been inoculated with GARDASIL® intramuscularly.

[000192] This tumor completely regressed and involuted soon after the first treatment,

with no further evidence of malignancy.

Example 6 - prostate cancer

[000193] Prostate cancer treatment would involve treating the patient with intramuscular

HPV, and can also include direct injection into the prostate.

Example 7 - glioblastoma multiforme

[000194] Glioblastoma multiforme treatment would involve treating the patient with

intramuscular HPV, and then direct injection into a tumor of glioblasotma multiforme.

PCT/US2021/054622

Example 8 - cervical cancer

[000195] Cervical cancer treatment would involve treating the patient with intramuscular

HPV, and can also include direct injection into the cervix.

Example 9 - anal cancer

[000196] Anal cancer treatment would involve treating the patient with intramuscular

HPV, and can also include direct injection or topical application to the anus.

Example 10 - Case Report: Use of vaccine plus N-Acetyl Cysteine (NAC)

[000197] The patient is a 74-year old woman who has an approximately 20 year history

of more than 70 keratinocyte carcinomas. The patient received the Gardasil vaccine in a series

of three doses. This course was repeated two years later.

[000198] The patient did not respond to treatment. There was no observed decrease in the

number of skin cancers per unit of time in this particular patient. These two courses of Gardasil

did not seem to decrease the number of skin cancers that she was having.

[000199] The patient was given a course of an mTOR inhibitor (sirolimus 10mg/week for

6 weeks) as per the protocol on this study.

[000200] The patient at this time was also started on N-acetylcysteine 600mg bid. When

the patient returned for her follow up two months later the patient had no visible skin cancers.

This observation was confirmed by physical examination.

[000201] In addition to the absence of keratinocyte carcinomas, the appearance of the

patient's skin was improved with the appearance of fewer actinic keratoses precancers also.

[000202] It is unknown whether a prolonged remission of skin cancers in this patient will

result. Follow up is scheduled.

[000203] Use of other HPV vaccines in treating cancer or tumors in accordance with the

methods described herein, are fully contemplated and are within the scope of the invention.

[000204] While the present invention has been presented in accordance with several

preferred and practical embodiments thereof, it is recognized that departures from the instant

disclosure are fully contemplated within the spirit and scope of the invention.

Claims

CLAIMS CLAIMS 29 Jul 2025 2021360676 29 Jul 2025

1. 1. A method A methodofoftreating treatingaa patient patient having skin cancer, having skin cancer, cancerous tumor,oror cancerous cancerous tumor, cancerouslesion, lesion, said said method comprising method comprising administering administering to to thethe patient,a apharmaceutical patient, pharmaceutical composition composition consisting consisting

of: of:

aa human papillomavirus human papillomavirus (HPV) (HPV) vaccine vaccine selected selected from from the group the group consisting consisting of an of HPVan HPV

quadrivalent recombinantvaccine quadrivalent recombinant vaccinecomprising comprising HPVHPV L1 proteins L1 proteins of types of HPV HPV 6,types 11, 6, 1611, and16 and 18, 18,

and an HPV HPVmultivalent multivalentrecombinant recombinant vaccine comprising HPV HPV L1 proteins of HPVoftypes HPV16, types 18,16, 18, 2021360676

and an vaccine comprising L1 proteins

31, 33, 45, 31, 33, 45,52, 52,and and58;58; a second a active pharmaceutical second active ingredient which pharmaceutical ingredient whichis is an an immune-based therapeuticselected immune-based therapeutic selected from an Immune from an Immune Checkpoint Checkpoint Inhibitor Inhibitor andand Chimeric Chimeric Antigen Antigen Receptor Receptor (CAR) (CAR) T-cells; T-cells; and and aa pharmaceutically acceptable pharmaceutically acceptable carrier. carrier.

2. 2. The method The methodofofclaim claim 1, 1, wherein wherein thethe Immune Immune Checkpoint Checkpoint Inhibitor Inhibitor is selected is selected from from the the group consisting of group consisting of aa Programed ProgramedDeath Death 1 inhibitor,a aProgramed 1 inhibitor, Programed Death Death Ligand Ligand inhibitor, inhibitor, and and a a Cytotoxic T-lymphocyte-Associated Cytotoxic T-lymphocyte-Associated Protein Protein 4 inhibitor. 4 inhibitor.

3. 3. The method The methodof ofclaim claim 2, 2, wherein wherein thethe Programed Programed DeathDeath 1 inhibitor 1 inhibitor or Programed or Programed Death Death Ligandinhibitor Ligand inhibitor isis Cemiplimab, Cemiplimab, Atezolizumab, Atezolizumab, Avelumab, Avelumab, Bavencio, Bavencio, Durvalumab, Durvalumab, Imfinzi, Imfinzi, Keytruda, Nivolumab, Keytruda, Nivolumab, Opdivo, Opdivo, Pembrolizumab, Pembrolizumab, or Tecentriq. or Tecentriq.

4. 4. The method The method of any of any one one of claims of claims 1 to 1 to 3, 3, wherein wherein the CARthe CARare T-cells T-cells are bispecific bispecific antibodies antibodies

or or monoclonal antibodies. monoclonal antibodies.

5. 5. The methodofofclaim The method claim4,4,wherein whereinthe thebispecific bispecificantibodies antibodiesare are Catumaxomab. Catumaxomab.

6. 6. The methodofofclaim The method claim 4, 4, wherein wherein the the monoclonal monoclonal antibodies antibodies are Campath, are Campath, Brutuximab, Brutuximab,

Vismodigib, Vemurafenib, Vismodigib, Vemurafenib, Dabrfenib,or Dabrfenib,or ecorafenib. ecorafenib.

7. 7. The method The method of any of any oneclaims one of of claims 1 to 6,1 wherein to 6, wherein the is the vaccine vaccine is substantially substantially free of host- free of host-

cell cell early antigen,E6E6ororE7.E7. early antigen,

8. 8. The method The methodofofanyany oneone of of claims claims 1 7, 1 to to 7, wherein wherein the the skinskin cancer, cancer, cancerous cancerous tumor, tumor, or or cancerous lesion cancerous lesion is is substantially substantially reduced reduced in or in size size or eliminated. eliminated.

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9. 9. The method of any one of claims 1 to 7, wherein the cancer or cancerous lesion is selected The method of any one of claims 1 to 7, wherein the cancer or cancerous lesion is selected 29 Jul 2025 2021360676 29 Jul 2025

from the group from the group consisting consisting of squamous cell of squamous cell carcinoma, carcinoma, basal basal cell cell carcinoma, carcinoma, melanoma, melanoma,

glandular tumor, and glandular tumor, and adenoma. adenoma.

10. 10. The The method method ofone of any anyofone of claims claims 1 to 1 9,towherein 9, wherein the patient the patient is 27 is 27 years years of of ageage or or older older oror

is is previously previously not not immunized withananHPV immunized with HPV vaccine. vaccine.

11. 11. TheThe method method of any of any one one of claims of claims 1 to110, to 10, wherein wherein the method the method further further comprises: comprises: 2021360676

establishing establishing a positive diagnosis a positive of skin diagnosis of skin cancer, cancer, diagnosis diagnosisofofbenign benignor or cancerous cancerous tumor, tumor, or or

diagnosis diagnosis ofof HPV HPV infection, infection, priorprior to administering to administering thedose the first firstofdose of the the HPV HPV vaccine. vaccine.

12. 12. The The method method ofone of any anyofone of claims claims 1 tosaid 1 to 11, 11, method said method comprising comprising administering administering a dose a dose of the pharmaceutical of the pharmaceuticalcomposition composition directly directly to a to a tumor tumor or skinorcancer skin lesion cancerorlesion or an area an area

immediately surroundingthe immediately surrounding thetumor, tumor,ororskin skincancer cancerlesion. lesion.

13. 13. The The method method of anyofone anyofone of claims claims 1 wherein 1 to 11, to 11, wherein the pharmaceutical the pharmaceutical composition composition is is administered administered by by injection. injection.

14. 14. The The method method of anyofone anyofone of claims claims 1 wherein 1 to 11, to 11, wherein the pharmaceutical the pharmaceutical composition composition is is administered administered to to thethe patient patient infixed-dose in a a fixed-dose combination combination product.product.

15. 15. The The method method of anyofone anyofone of claims claims 1 towherein 1 to 14, 14, wherein the patient the patient is administered is administered a second a second

dose of vaccine dose of vaccine and andimmune-based immune-based therapeuticat atleast therapeutic leastoneone month month following following the the first first

administration, andoptionally administration, and optionallyisisadministered administered a third a third dosedose of vaccine of vaccine and immune-based and immune-based

therapeutic at therapeutic at least leastone onemonth month following following the the second administration. second administration.

16. 16. An anticancer An anticancer pharmaceutical pharmaceutical composition composition consisting consisting of: of: an an HPV vaccineselected HPV vaccine selectedfrom from thegroup the group consistingofofananHPV consisting HPV quadrivalent quadrivalent recombinant recombinant

vaccine comprising vaccine comprisingHPV HPV L1 protein L1 protein of HPV of HPV types types 6, 11,6,16, 11,and 16,18, andand 18,anand HPVan HPV multivalent multivalent

recombinant vaccinecomprising recombinant vaccine comprisingHPVHPV L1 protein L1 protein of HPV of HPV types types 16,31, 16, 18, 18,33, 31, 45, 33, 52, 45, 52, and and 58; 58;

a second a active pharmaceutical second active ingredient which pharmaceutical ingredient whichis is an an immune-based therapeuticselected immune-based therapeutic selected from an Immune from an Immune Checkpoint Checkpoint Inhibitor Inhibitor andand CAR CAR T-cells; T-cells; and and

aa pharmaceutically acceptable pharmaceutically acceptable carrier. carrier.

17. 17. The composition The compositionofofclaim claim16, 16,wherein whereinthe theImmune Immune Checkpoint Checkpoint Inhibitor Inhibitor is selected is selected from from

the group the consisting of group consisting of aa Programed Death1 1inhibitor, Programed Death inhibitor, aa Programed Death Programed Death Ligand Ligand inhibitor,and inhibitor, and aa Cytotoxic T-lymphocyte-Associated Cytotoxic T-lymphocyte-Associated Protein Protein 4 inhibitor. 4 inhibitor.

46

18. 18. The composition The compositionofofclaim claim 17,17, wherein wherein thethe Programed Programed DeathDeath 1 inhibitor 1 inhibitor or Programed or Programed 29 Jul 2025 2021360676 29 Jul 2025

Death Ligand inhibitor Death Ligand inhibitor is is Cemiplimab, Atezolizumab, Avelumab, Cemiplimab, Atezolizumab, Avelumab,Bavencio, Bavencio,Durvalumab, Durvalumab, Imfinzi, Imfinzi, Keytruda, Nivolumab,Opdivo, Keytruda, Nivolumab, Opdivo, Pembrolizumab, Pembrolizumab, or Tecentriq. or Tecentriq.

19. 19. The composition The compositionofofany anyone one ofof claims1616 claims to to 18,wherein 18, wherein thethe CAR CAR T-cells T-cells are are bispecific bispecific

antibodies antibodies which is Catumaxomab which is Catumaxomab or or monoclonal monoclonal antibodies antibodies Campath, Campath, Brutuximab, Brutuximab, Vismodigib, Vismodigib,

Vemurafenib, Dabrfenib,ororecorafenib. Vemurafenib, Dabrfenib, ecorafenib. 2021360676

47