AU2017223970B2 - Method and composition for treating cancer or skin lesion using a vaccine - Google Patents

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 one or more doses of HPV recombinant vaccine to a patient.

Description

METHOD AND COMPOSITION FOR TREATING CANCER OR SKIN LESION USING A VACCINE FIELDOFTHEINVENTION

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

malignant tumor and, more particularly, to a method for treatment, or reducing the incidence of

recurrence, of cancer or tumors comprising administration of a vaccine, including local

administration of a composition comprising the vaccine as a therapeutic agent.

BACKGROUND OF7THE INVENTION

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

squamous cell carcinoma (SCQ and melanoma, and 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). HWV 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 -PV18. HPV is not known to be a cause or to be associated with basal

cell carcinoma (13CC) or melanoma.

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

and other conditions caused by or associated with IPV 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 HV (types 16, 18, 31, 33, 45, 52, and 58). GARDASIL© is indicated for use in

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

and in boys from ages 9-15.

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

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

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

HPV (as in GARDASIL*), and additional 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 require more than viral-specific components, such as HPV LI proteins that comprise the commercially available preventive vaccines, such as 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 requires Trojan 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 [7 would not be expected to work on cells that do not have E6 orE7,

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] It is an object of the present invention to overcome or ameliorate at least one

of the disadvantages of the prior art, or to provide a useful alternative. 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.

[00014] Any discussion of the prior art throughout the specification should in no way

be considered as an admission that such prior art is widely known or forms part of common

general knowledge in the field

[00014a] Unless the context clearly requires otherwise, throughout the description and

the claims, the words "comprise", "comprising", and the like are to be construed in an

inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of

"including, but not limited to".

SUMMARY OF THE INVENTION

[00014b] The subject invention concerns a method for treating a patient having skin

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

papilloma virus (HPV) infection, or other skin lesion, said method comprising the steps of:

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

other skin lesion, a therapeutically effective dose of a commercially available HPV vaccine.

The vaccine can be administered directly to the cancer 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.

[00014c] In another aspect, the present disclosure provides a method of treating squamous

cell carcinoma in a subject the method comprising administering to the subject a

therapeutically effective amount of a pharmaceutical composition comprising:

at least one purified viral LI protein, wherein the at least one purified viral LI is a

human papilloma virus (HPV) Li protein and is free of host-cell and/or non-L I HPV peptide,

a second active pharmaceutical ingredient selected from the group consisting of a local

anesthetic, sirolimus, ingenol mebutate and T4 endonuclease; and

a pharmaceutically-acceptable carrier,

wherein the pharmaceutical composition comprises a purified viral LI protein or

fragment thereof from each of HPV types 6, 11, 16 and 18,

wherein the pharmaceutical composition is administered by injection.

[00014d] In another aspect, the present disclosure provides use of:

at least one purified viral Li protein, wherein the at least one purified viral LI

is a human papilloma virus (HPV) LI protein and is free of host-cell and/or non-Li HPV

peptide,

a second active pharmaceutical ingredient selected from the group consisting of

a local anesthetic, sirolimus, ingenol mebutate and T4 endonuclease; and

a pharmaceutically-acceptable carrier,

in the manufacture of a medicament for the treatment of squamous cell

carcinoma, wherein the medicament is administered by injection,

wherein the medicament comprises a purified viral LI protein or fragment

thereof from each of HPV types 6, 11, 16 and 18.

[00014e] 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 thefirst administration; and

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

about five months to about seven months after the first dose

5a

[00015] 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.

[00016] 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.

[00017] The -V vaccine can be selected from HPV quadrivalent (types 6, 11, 16, and

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

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

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

[00018] In one preferred embodiment, the method does not comprise or is without

administering an additional or other immunostimulant or adjuvant.

[00019] In one preferred embodiment, the method comprises administering an additional

or other immunomodulatory agent, such as an immunostimulant or adjuvant.

[00020] 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.

[00021] 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.

[00022] 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.

[00023] Each dose of HPV vaccine administered in the above method steps is preferably

about 0.5 ml, and is more preferably 0.5 ml.

[00024] The method can further comprise establishing a positive diagnosis of cancer,

benign tumor, or HPV infection prior to administering the first dose ofHPV vaccine.

[00025] An alternative embodiment of the method according to the subject invention

comprises treating a patient having cancer, benign tumor, or ahuman papilloma virus-related

(I-IPV-related) 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.

[00026] 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.

[00027] These direct second or third administrations of a composition comprising the

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

[00028] 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.

[00029] 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.

[00030] The preferred dose of each subsequent administration of HPV vaccine, if any, is

0.5 ml.

[00031] The method according to any embodiment of the invention can be used for

treating cancer, benign tumor, or HPV-related lesion, including, but not limited to, a benign tumor associated or unassociated with HPV infection, squamous cell carcinoma, basal cell carcinoma, melanoma, verruca vulgaris, and condyloma accuminata.

[00032] The method can further comprise establishing a positive diagnosis of cancer,

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

[00033] 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.

[00034] 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.

[00035] 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.

[00036] 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.

[00037] 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.

[00038] 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.

[00039] Compositions comprising the vaccine are also included as part of the invention.

For example, the 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.

[00040] 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 to 10:1 (v/v) vaccine:anesthetic solution can be used, as would be

understood in the art.

[00041] 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.

[00042] Solutions for injection can also include one or more buffer, emollient, diluent, p1H

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

[00043] 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.

[00044] 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.

[00045] 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.

[00046] To the knowledge of the inventor, administration of HPV vaccines comprising

only HPV antigens (being free of host-cell peptides), to a previously unimmunized patient, or an

adult patient aged 27 or greater, to eliminate or reduce the incidence of recurrence of skin cancer,.

benign or malignant tumor or other skin lesion that is not an -IPV-associated lesion, has not been

previously described. Nor has the direct or local administration of a vaccine by topical

application or by direct injection into the lesion or tumor been previously described to eliminate

the lesion and reduce the incidence of its recurrence.

DETAILED DESCRIPTION

[00047] The present invention is directed to a method of treating cancer, benign tumor,

skin cancer, such as squamous cell carcinoma (SCC), or a 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, to a patient having a cancer or tumor.

[00048] 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.

[00049] 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.

[00050] 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.

[00051] 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.

[00052] 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.

[00053] 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.

[00054] 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 (El-E2) and two in the "late" region (LI (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.).

[00055] In one embodiment, an HPV vaccine of the subject invention comprises one or

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

nine, ten or more different IPV 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.

[00056] In one embodiment, the HPV vaccine employed in the subject method contains

purified inactiveviral or virs-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-LI HPV peptide,

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

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

[00057] 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.

[00058] Alternatively, the vaccine can be administered to reduce the incidence of

recurrence of cancer, a benign tumor, or an HPV-related lesion in an unimmunized patient. 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 an adult patient aged 27 or greater.

[00059] More particularly, one preferred embodiment of the invention comprises a method

for the treatment of cancer, benign tumor or IPV-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. 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.

[00060] 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.

[00061] 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 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 abouttwo 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.

[00062] 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.

[00063] 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, osteochronfroma (osteocartilaginous exostoses), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (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.

[00064] 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, adenoids

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

cancer includes liver cancer.

[00065] 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

(BAKI). 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 BAKI 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.

[00066] 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.

[00067] 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.

[00068] 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.

[00069] Other particular types of cancers or tumors that can benefit from treatment using

an [PV 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.

[00070] 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., verrucavulgaris or condyloma accuminata

[00071] 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.

[00072] 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.

[00073] A phannaceutical 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), transdennal (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, glycerine, 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

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.

[00074] 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 ELTM (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 syringability 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, orby 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.

[00075] 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.

[00076] 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

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.

[00077] 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.

[00078] 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.

[00079] 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.

[00080] 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 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 US. Pat. No. 4,522,811.

[00081] It is especially advantageous to formulate oral or parenteral compositions in

dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as 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 are 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.

[00082] Toxicity and therapeutic efficacy of such compounds can be determined by

standard pharmaceutical procedures in cell cultures or experimental animals, eg., 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.

[00083] 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.

[00084] The pharmaceutical compositions can be included in a container, pack or

dispenser together with optional instructions for administration.

[00085] 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

compositions 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.

[00086] 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, or to specific strata

thereof, or to an underlying tissue.

[00087] 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.

[00088] 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 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.

[00089] 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.

[00090] The types of pharmaceutical excipients that are useful as carriers include

stabilizers such as Human Serum Albumin (ISA), 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.

[00091] 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 as2-hydroxypropyl-p-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.

[00092] 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.

[00093] 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 that the drug

can be applied, localized and removed easily.

[00094] 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 IPV, such as warts, e.g., verruca vulgaris or condyloma accuminata.

[00095] 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.

[00096] 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.

[00097] 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.

[00098] 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. 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

[00099] 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.

[000100] 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.

[000101] 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, trietylenephosphoramide,

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 I 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, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, 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 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 anti-hormonal agents that act to regulate or inhibit hormone action on tumors such as anti-estrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen, raoxifene, droloxifene, 4-hydroxytamoxifen, trioxifene, keoxifene,

LYI17018, 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.

[000102] 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) which comprises the therapeutic agent and one or more pharmaceutically

acceptable carriers, excipients and diluents, according to standard pharmaceutical practice.

[000103] Each therapeutic agent in a combination therapy of the invention may be

administered simultaneously (ie., 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.

[000104] 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.

[000105] 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.

[000106] 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.

[000107] 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.

[000108] 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.

[000109] 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.

[000110] Any commercially available HPV vaccine can be employed for administration

directly to a cancer orI-IPV-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 HPV

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

bivalent (types 16 and 18) recombinant vaccine.

[000111] 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 patientorapatient previously

immunized with an -IPV vaccine.

[000112] 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.

[000113] In another embodiment, the vaccine can be administered to treat cancer, benign

tumor, or an 1-PV-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.

[000114] More particularly, one preferred embodiment of the invention comprises a method

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

[000115] 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 HIV

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

[000116] 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 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.

[000117] 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 are 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.

[000118] 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 atleast 0.05 ig/kg, 0.2 jg/kg, 0.5 pg/kg, 1 ig/kg, 10 pg/kg, 100 pg/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.

[000119] 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

days ( 2 days) or about 30 days (± 2 days) or about one week (± 2 days), two weeks ( 2 days),

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

[000120] In other embodiments, the dosing regimen will comprise administering the HPV

vaccine at a dose of from about 0.005 mg/kg to about 10mg/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 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 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.

[000121] 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 HIV 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 HIPV 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.

[000122] 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 -V 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.

[000123] In one exemplary embodiment, each secondary and/or tertiary dose is

administered Ito 14 (e.g., 1, 1%,2, 2 !3,3%,4,4%,5,5%,/6,6 %,7, 7½,8,8%,9,9%, 10, 10,

11,11½,., 12, 12%, 13, 13, 14, 14 , or more) weeks after the immediately preceding dose. In

another exemplary embodiment, each secondary and/or tertiary dose is administered I to 14

(e.g., 1, 1,2,2,3,3%, 4, 4½,5,5%,6,6%, 7, 7, 8, 8%,9,9%, 10, 10%, 11, 11%, 12, 12½,

13, 13%, 14, 14½, or more) months after the immediately preceding dose. The phrase "the

immediately preceding dose," as used herein, means, in a sequence of multiple 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.

[000124] 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.

[000125] 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 I 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 I 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.

[000126] In certain embodiments, the initial dose (eg., 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.

[000127] 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.

[000128] 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.

[000129] 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

associated, but non-cancerous, lesion, such as warts, including genital warts, e.g., verruca

vulgaris or condyloma accuminata.

[000130] 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

HIV-associated, but non-cancerous, lesion, such as warts, including genital warts, e.g., verruca

vulgaris or condyloma accuminata.

[000131] 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

vaccine.

[000132] 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 H1V infection, such as basal cell carcinoma or

melanoma.

[000133] 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.

[000134] 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

[000135] 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.

[000136] Topical application can be beneficial for several reasons, including the

elimination of infection risk caused by injection, but can also be advantageous by wide-spread

application over large areas in order to treat precancerous (actinic keratoses) as well as

malignant tumors. In addition, the topical administration can provide cosmetic enhancement

of the skin, by decreasing that appearance of pigment irregularities, poikiloderma, and scaling.

[000137] The subject invention relates to a cost-effective, safe, efficacious, and

convenient treatment for reducing or ameliorating the growth or size of a cancer tumor or

lesion, including a skin cancer lesion such as SCC, BCC or melanoma tumor or lesion. The

subject invention also relates to a cost-effective, efficacious and convenient treatment for

curing skin cancer lesions, and further relates to a cost-effective, efficacious and convenient

method to reduce the incidence of recurrence of cancer, including skin cancer lesions.

[000138] The subject method of treating or reducing the incidence of recurrence of skin

cancer comprises administering an HPV vaccine in one or more doses to a patient. In one

embodiment, the method includes administration of a first dose of HPV quadrivalent (types 6,

11, 16, and 18) recombinant vaccine to a patient, a second dose of HPV quadrivalent (types 6,

11, 16, and 18) recombinant vaccine approximately two months thereafter, and a third dose of

HPV quadrivalent (types 6, 11, 16, and 18) recombinant vaccine approximately four months after

the second dose. In a preferred embodiment, each dose is 0.5 ml.

[000139] The subject method can be advantageous in that it can be performed using a

commercially available HIPV bivalent (types 16 and 18) recombinant vaccine, HPV quadrivalent

(types 6, 11, 16, and 18) vaccine or HPV multivalent (types 16, 18, 31, 33, 45, 52, and 58)

recombinant vaccine as a therapeutic agent rather than or in addition to its use as a preventive

vaccine.

[000140] 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 HPV quadrivalent

(types 6, 11, 16, and 18) recombinant vaccine and GARDASIL* 9, is an 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).

[000141] By use of a commercially available vaccine, the vaccine can be readily accessed

by a physician or healthcare practitioner. Moreover, the use of an -IPV bivalent (types 16 and

18) recombinant vaccine, an -V 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-L viral peptides, polypeptides, or proteins,

such as the antigens, E6 or E7.

[000142] 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 -IPV infection, can be achieved using the subject method as

described herein.

[000143] 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 ingredient, excipient or diluent, 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.

[000144] 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; and 7) T4 endonuclease 8) Antimetabolites, e.g. 5 Fluorouracil, Methotrexate 9) cyclooxygenase inhibitors, e.g. Diclofenac

[000145] 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.

[000146] 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.

[000147] 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 to 10:1 (v/v) vaccine:anesthetic solution can be used, as would be understood in the art.

[000148] 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 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.

[000149] Solutions for injection can also include one or more buffer, emollient, diluent, pH

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

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.

[000150] 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.

[000151] 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.

[000152] 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

EXAMPLES

Example I - skin cancer

[000153] 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.

[000154] 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

[000155] Patient I 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 I 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 I had more than 300 distinctive occurrences of

skin cancer during his lifetime.

PATIENT I Time Period SCC BCC (Months) Prior to Commencement 16 1.80 0.25 of Treatment Method After Commencement of 16 0.37 0.00 Treatment Method

B. Patient 2

[000156] 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 BCC

Prior to Commencement 13onths5 13 2.07 0.53 of Treatment Method After Commencement of 13 0.23 0.3 Treatment Method

C_ Patient3

[000157] 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 0.18 013 of Treatment Method After Commencement of 22 0.09 0.04 Treatment Method

[000158] 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.

[000159] 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.

[000160] In one embodiment, the method of treatment for eliminating or reducing the

incidence of recurrence of skin cancer includes administering the 1-PV 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

[000161] 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.

[000162] 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 -IPV 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.

[000163] 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

radius cubed).

[000164] With the expected doubling of size, the tumor should have increased by about

40%. to a tumor diameter of about 4.6 centimeters.

Example 3 - metastatic ba.sosquanou carcinoma

[000165] 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.

[000166] 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.

[000167] 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

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 was treated with

three equal doses of GARDASIL*, intramuscularly, in accordance with the label instructions.

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.

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 --- aggressivesquamous cell carcinoma

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.

The patient had previously been inoculated with GARDASIL* intramuscularly.

This tumor completely regressed and involuted soon after the first treatment, with no further

evidence of malignancy.

Example 6 -prostate cancer

Prostate cancer treatment would involve treating the patient with intramuscular HPV, and can

also include direct injection into the prostate.

Example 7 - glioblastomamultiforme

Glioblastoma multiforme treatment would involve treating the patient with intramuscular HPV,

and then direct injection into a tumor of glioblasotma multiforme.

Example 8 --- cervical cancer

Cervical cancer treatment would involve treating the patient with intramuscular HPV, and can

also include direct injection into the cervix.

Example 9 - anal cancer

[000168] Anal cancer treatment would involve treating the patient with intramuscular HPV,

and can also include direct injection or topical application to the anus.

[000169] 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.

[000170] 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 (10)

1. Claim 1. A method of treating squamous cell carcinoma in a subject the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising: at least one purified viral Li protein, wherein the at least one purified viral Li is a human papilloma virus (HPV) Li protein and is free of host-cell and/or non-L HPV peptide, a second active pharmaceutical ingredient selected from the group consisting of a local anesthetic, sirolimus, ingenol mebutate and T4 endonuclease; and a pharmaceutically-acceptable carrier, wherein the pharmaceutical composition comprises a purified viral Li protein or fragment thereof from each of HPV types 6, 11, 16 and 18, and wherein the pharmaceutical composition is administered by injection.
2. Claim 2. The method of claim 1, wherein the pharmaceutical composition comprises a purified viral Li protein or fragment thereof from each of HPV types 16, 18, 31, 33, 45, 52 and 58.
3. Claim 3. The method of any one of claim I or claim 2, wherein the at least one purified viral Li protein is present in a virus-like particle (VLP).
4. Claim 4. The method of any one of claims I to 3, wherein the at least one purified viral Li protein and the second active pharmaceutical ingredient and pharmaceutically acceptable carrier are in a unitary dosage form.
5. Claim 5. The method of any one of claims I to 4, wherein the local anesthetic is lidocaine.
6. Claim 6. Use of: at least one purified viral Li protein, wherein the at least one purified viral L is a human papilloma virus (HPV) Li protein and is free of host-cell and/or non-Li HPV peptide, a second active pharmaceutical ingredient selected from the group consisting of a local anesthetic, sirolimus, ingenol mebutate and T4 endonuclease; and a pharmaceutically-acceptable carrier, in the manufacture of a medicament for the treatment of squamous cell carcinoma, wherein the medicament is administered by injection, wherein the medicament comprises a purified viral Li protein or fragment thereof from each of HPV types 6, 11, 16 and 18.
7. Claim 7. The use of claim 6, wherein the medicament comprises a purified viral LI protein or fragment thereof from each of HPV types 16, 18, 31, 33, 45, 52 and 58.
8. Claim 8. The use of claim 6 or claim 7, wherein the at least one purified viral LI protein is present in a virus-like particle (VLP).
9. Claim 9. The use of any one of claims 6 to 8, wherein the at least one purified viral LI protein and the second active pharmaceutical ingredient and pharmaceutically acceptable carrier are in a unitary dosage form.
10. Claim 10. The use of any one of claims 6 to 9, wherein the local anesthetic is lidocaine.