A Phase 1 interventional study of 5-fluorouracil and Calcipotriene in Carcinoma, Squamous Cell, sponsored by Melissa Pugliano-Mauro. Recruiting at 1 site in United States. Open to participants aged 18 Years and older, including healthy volunteers. Per ClinicalTrials.gov, last updated 2026-06-01.
Sponsored by Melissa Pugliano-Mauro · Phase 1, Interventional, and Treatment
This clinical trial proposes to evaluate a relatively unexplored approach to treatment of squamous cell carcinoma (SCC) on the lower extremities. The strategy is to directly and specifically deliver drug to the tumor. For the proposed phase I clinical trial, the investigators will perform intralesional injections of a well characterized, potent chemotherapeutic agent 5-fluorouracil (5FU) with and without a topical application of 0.005% calcipotriene cream to kill topically accessible SCC cells. The goal of the study is to evaluate the safety profile and tolerability of intralesional-5FU with and without a concomitant topical calcipotriene and measure the clinical objective response rate (ORR) in treated lesions compared to untreated lesions 3 weeks after treatment.
Squamous cell carcinoma of the skin is a common form of keratinocyte skin cancer. The majority of cutaneous SCCs occur on the head and neck, more so in men than women, but can occur anywhere squamous cells are found. Squamous cell carcinoma of the skin is usually not life-threatening, though it can be aggressive in some cases, and is normally treated with minor surgery.
Squamous cell carcinoma (SCC) of the lower extremity is a distinct subset of cutaneous squamous cell carcinomas which tend to occur multiply in elderly women. Histopathological studies of lower extremity SCCs reveals that they tend to be well differentiated and have low incidence of perineural and lymphovascular invasion and are also less prone to metastasis. Minor surgery has been the standard of care for this subset of SCC but leads to many complications such as poor wound healing and postoperative infections. Furthermore, a phenomenon called eruptive postoperative SCC can occur, in which cytokines released during wound healing triggering secondary tumor formation in genetically predisposed cells surrounding the original SCC. Given that lower extremity SCCs are less aggressive but more prone to surgical complications when excised, we believe they may be good candidates for localized non-surgical treatment.
5-fluorouracil (5FU) is a well-studied and characterized chemotherapeutic agent that has been used systemically, topically and intralesionally for a variety of malignancies and conditions including many dermatologic abnormalities.
It has been used on and off label topically for the treatment of actinic keratosis, SCC, superficial basal cell carcinoma, extra-mammary Paget's disease confined to the epidermis, Bowen's disease, porokeratosis, and genital warts. Intralesionally it has been used off label for the treatment of keloids, knuckle pads, warts, hypertrophic scars, basal cell carcinoma, and keratoanthoma.
Reports of its use intralesionally in invasive cutaneous SCC, other than in keratoacanthomas, are very limited. We are aware of 3 such reports in the literature. In the largest study to date, 6 weekly intralesional injections of 5FU-epinephrine gel were performed on 23 patients with cutaneous SCC on various body sites, 22 (96%) of whom demonstrated histologically confirmed tumor clearance. This study, however, used a proprietary gel formulation which is not widely available. There are two other case reports of successful treatment of SCC with 6-8 intralesional injections of 5FU at weekly intervals. The three published studies injected 0.6ml to 2.4ml of 5FU, per each weekly session, at concentrations of 30mg/ml to 50mg/ml.
Calcipotriene is a synthetic form of vitamin D. It is FDA approved for the treatment of psoriasis. Calcipotriene binds to vitamin D receptors on skin cells and helps regulate the growth and differentiation of skin cells. It inhibits keratinocyte proliferation (cell growth) and enhances keratinocyte differentiation. Recently it has been shown that calcipotriene acts as topical immune response modulator through induction of thymic stromal lymphoprotein (TSLP) expression. TSLP is an epithelium-derived cytokine and a regulator of allergic inflammation in the skin. It has been shown by Shadmehr Demehri et.al. that TSLP released by barrier-defective skin in mice blocks cancer development by recruiting T cells to mount robust antitumor immunity in the skin. The adaptive immune response mounted by TSLP against cancer can eliminate cancerous lesions in the skin and prevent new lesions from developing.
A recent clinical trial conducted by Demehri showed that participants treated with topical 5FU and topical and topical calcipotriene showed a significant reduction in the number of actinic keratosis (face, scalp, \& upper extremities) vs topical 5FU and vaseline. Furthermore, the cohort treated with 5FU and topical and topical calcipotriene remained SCC-free for more than 1,500 days suggesting the induction of tissue-resident memory T (trm) cells.
The purpose of the study is to evaluate this relatively unexplored approach to the treatment of SCC on the lower extremities and establish a non-surgical therapy to improve outcomes, eliminate the need for surgery and mitigate the occurrence of infections and secondary tumor eruptions.
Topical 5FU is approved for the treatment of superficial basal cell carcinoma smaller than 2cm in diameter and not located on the feet, hands or feet but cannot penetrate deep enough into the skin to be an effective treatment for SCC. For the proposed phase I clinical trial, we propose to increase tumor accessibility by delivering 5FU directly and specifically to the tumor through intralesional injections to kill accessible localized SCC cells.
5FU is currently in clinical use with a well-established safety profile. It is anticipated that intralesional injections of 5FU will enable direct and specific delivery of chemotherapy to the tumor, thereby reducing the potential for systemic toxicity. Further, intralesional injections of-5FU enable tumoral delivery of locally effective concentrations of 5FU using doses that are orders of magnitude below those used currently for the intravenous (IV) treatment of multiple malignancies.
5FU alone may not be enough eliminate all the tumor cells so we also propose to treat a group of patients with intralesional 5FU and 0.005% calcipotriene topical cream. The goal is to develop a synergistic effect and establish an immune response to the 5FU induced apoptotic SCC cells in order to kill off any remaining SCC cells not undergoing apoptosis. Also 5FU alone may not be as effective of a treatment as the combination therapy in the prevention of secondary tumor recurrence. It is hoped that the combination therapy will induce a memory response and reduce the incidence of secondary tumors.
Participants will have at least 1 SCC lesion greater 1cm and less than 2 cm in largest diameter, on their lower extremities. The clinical diagnosis of SCC will be confirmed histologically by a deep shave biopsy of less than half of the lesion. The remainder of the lesion will be used for intralesional injections of 5FU or intralesional 5FU/topical calcipotriene according to the following schema:
In this study, a total of 30 patients will be randomly assigned into 3 groups. Randomization will be conducted using the UPCI randomizer, which is maintained by the Biostatistics Facility of UPCI (https://randomize.upci.pitt.edu/randomizer/home.seam). Group 1: 10 patients will serve as a control group, and will receive neither 5FU injection nor topical calcipotriene. Group 2: 10 patients, will receive a single intralesional injection of 50mg of 5FU in 1ml aqueous injectable solution once a month for 3 months. Group 3: 10 patients will receive intralesional 5FU as administered in the previous group. They will also apply a topical application of 0.005% calcipotriene cream to the same lesion two (2) times daily for four (4) days one (1) day after each of three (3) 5FU injections. At the time of initial consent, a 2 mm punch biopsy of the lesion will be obtained. Half of biopsy will be stored for tissue banking and future study, while the other half will be sent for histological confirmation of SCC. A second 2 mm punch biopsy will be taken at 2 months for mid-point analysis. This will be stored for tissue banking and future study. A month after the last injection (week 12), the lesion will be surgical resected in the non-treated Group 1 to render the participant's disease free. Resection is the current standard of care for these tumors. Participant's in Group 2 and 3 will have a 4mm punch biopsy will be taken to confirm pathologic resolution.
Depending on the results of the confirmational biopsy the following actions will be taken:
All lesions will be photographed and treatment response will be evaluated 4 weeks after the first 5FU injection prior to excision.
627 studies on the registry are indexed under Carcinoma, Squamous Cell; 133 are open to participants now.
This study's planned enrollment of 30 is below the median of 45 across 476 interventional studies indexed under Carcinoma, Squamous Cell.
Browse Carcinoma, Squamous Cell studies →Melissa Pugliano-Mauro is the lead sponsor of 3 studies on the registry; 3 are open to participants now.
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Go through the criteria one by one — yes, no or not sure — and take the list to your doctor or the study team. Kept in this browser only.
Exclusion Criteria:
Patients with the following cardiac co-morbidities including:
Patients with a known history of a hematologic , primary brain tumor or sarcoma, or of another primary solid tumor, unless the patient has undergone potentially curative therapy with no evidence of that disease for five years.
° The time requirement also does not apply to patients who underwent successful definitive resection of basal or squamous cell carcinoma of the skin, superficial bladder cancer, in situ cancers including cervical cancer, breast cancer, melanoma, or other in situ cancers.
Underlined terms in the criteria open these too. Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
A biopsy takes a small piece of tissue to examine. “Archival tissue” is a sample already taken, for example at diagnosis, and kept by the hospital.
Whether a stored sample will do depends on when it was taken, how much there is and what the study needs. Ask whether the study accepts your stored sample or needs a new biopsy.
A drug or device being studied in a clinical trial.
It may be new, or already approved and being studied for another use, dose or form. Criteria often leave out people who took part in another study of one within a recent period, commonly 30 days or five half-lives.
Before you join, the study team explains what taking part involves — its purpose, what happens, the risks and possible benefits — and answers your questions, so you can decide freely. Consent is usually recorded on a form (the informed consent form, ICF) signed by you or, where the law allows, by someone legally authorised to decide for you.
You can keep asking questions during the study, you are told of new information that matters, and you can choose to leave at any time.
A scale of how much an illness limits everyday activity. For people being assessed to join a study, scores run from 0 (fully active) to 4; lower is more active.
Studies most often ask for 0–1 or 0–2. Your doctor gives the score; the WHO scale uses the same steps. The full scale also has 5, which records death.
Able to become pregnant: from the first period until menopause, unless permanently unable to through surgery (removal of the womb, or of both ovaries or both fallopian tubes).
Criteria for people of childbearing potential usually ask for a negative pregnancy test and for contraception during the study and for a time after it.
Ways of preventing pregnancy. “Highly effective” methods fail less than 1% of the time a year when used correctly — for example an IUD or an implant.
Criteria usually say which methods count and how long to keep using them after the last dose, sometimes for male participants too. Some studies accept a partner’s vasectomy if it was confirmed successful and that person is the participant’s only sexual partner.
Cancer that has spread from where it began to other parts of the body. “Metastatic” cancer is cancer that has spread this way.
A weakened immune system. It can come from an illness, such as HIV or some blood cancers, or from medicines that turn the immune system down, for example after an organ transplant or for an autoimmune disease; steroids at higher doses count.
Read whether the line is about a condition or about medicines, and which medicines and doses it means.
Anti-inflammatory medicines such as prednisone, prednisolone and dexamethasone — not the anabolic steroids used for muscle.
Studies differ in what they allow: check which steroid medicines, routes (tablets, injections, creams, inhalers, eye drops) and doses this study accepts.
Hepatitis B is a virus that infects the liver. These are separate tests: HBsAg positive usually means a current infection (a recent vaccination can briefly cause it too); total anti-HBc (HBcAb) positive means an infection now or in the past; HBV DNA measures how much virus is in the blood.
Some studies accept a past infection if HBV DNA is undetectable, sometimes with antiviral medicine during the study. Check what this study allows.
The human immunodeficiency virus, which weakens the immune system.
Many studies now accept people whose HIV is controlled on treatment, judged by the viral load and the CD4 count. Check the limits this study sets.
A condition in which the immune system attacks the body’s own tissues, such as rheumatoid arthritis, lupus, psoriasis, type 1 diabetes or inflammatory bowel disease.
Immunotherapy studies often leave out active autoimmune disease, and some name exceptions, such as thyroid disease on replacement hormone. Check the exceptions this study lists.
On an ECG, the QT interval is the time the heart’s lower chambers take to activate electrically and recover in each beat. QTc is that time adjusted for heart rate (QTcF by Fridericia’s formula, QTcB by Bazett’s). Some medicines lengthen it, and a long QTc can raise the risk of a dangerous heart rhythm.
The study may name a correction formula and a limit in milliseconds, often around 450–480.
The share of blood the heart’s main pumping chamber (the left ventricle) pushes out with each beat, measured by an echocardiogram (heart ultrasound) or a MUGA scan. About 50–70% is normal.
The US National Cancer Institute’s scale of how severe an adverse event is — any unwanted medical event during a study, such as a symptom, an illness or an abnormal test result, whether or not a treatment caused it. Whether it did is judged separately.
“Side effects of earlier treatment resolved to grade 1 or less” means they have settled to mild or gone.
Malignancy means cancer. Criteria about another or earlier malignancy ask whether you have had a different cancer, usually within a set number of years.
Some are commonly allowed, such as treated skin cancers other than melanoma and early cancers removed completely — check the exceptions this study lists.
An allergy, or another reaction of the immune system, to a substance. “Hypersensitivity to the study drug” means a known allergic reaction to it — not every side effect a medicine can have.
Biopsy and archival tissue
A biopsy takes a small piece of tissue to examine. “Archival tissue” is a sample already taken, for example at diagnosis, and kept by the hospital.
Whether a stored sample will do depends on when it was taken, how much there is and what the study needs. Ask whether the study accepts your stored sample or needs a new biopsy.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Investigational drug or device
A drug or device being studied in a clinical trial.
It may be new, or already approved and being studied for another use, dose or form. Criteria often leave out people who took part in another study of one within a recent period, commonly 30 days or five half-lives.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Informed consent
Before you join, the study team explains what taking part involves — its purpose, what happens, the risks and possible benefits — and answers your questions, so you can decide freely. Consent is usually recorded on a form (the informed consent form, ICF) signed by you or, where the law allows, by someone legally authorised to decide for you.
You can keep asking questions during the study, you are told of new information that matters, and you can choose to leave at any time.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
ECOG performance status
A scale of how much an illness limits everyday activity. For people being assessed to join a study, scores run from 0 (fully active) to 4; lower is more active.
Studies most often ask for 0–1 or 0–2. Your doctor gives the score; the WHO scale uses the same steps. The full scale also has 5, which records death.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Childbearing potential
Able to become pregnant: from the first period until menopause, unless permanently unable to through surgery (removal of the womb, or of both ovaries or both fallopian tubes).
Criteria for people of childbearing potential usually ask for a negative pregnancy test and for contraception during the study and for a time after it.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Contraception
Ways of preventing pregnancy. “Highly effective” methods fail less than 1% of the time a year when used correctly — for example an IUD or an implant.
Criteria usually say which methods count and how long to keep using them after the last dose, sometimes for male participants too. Some studies accept a partner’s vasectomy if it was confirmed successful and that person is the participant’s only sexual partner.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Metastases
Cancer that has spread from where it began to other parts of the body. “Metastatic” cancer is cancer that has spread this way.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Immunosuppression
A weakened immune system. It can come from an illness, such as HIV or some blood cancers, or from medicines that turn the immune system down, for example after an organ transplant or for an autoimmune disease; steroids at higher doses count.
Read whether the line is about a condition or about medicines, and which medicines and doses it means.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Corticosteroids (steroids)
Anti-inflammatory medicines such as prednisone, prednisolone and dexamethasone — not the anabolic steroids used for muscle.
Studies differ in what they allow: check which steroid medicines, routes (tablets, injections, creams, inhalers, eye drops) and doses this study accepts.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Hepatitis B tests
Hepatitis B is a virus that infects the liver. These are separate tests: HBsAg positive usually means a current infection (a recent vaccination can briefly cause it too); total anti-HBc (HBcAb) positive means an infection now or in the past; HBV DNA measures how much virus is in the blood.
Some studies accept a past infection if HBV DNA is undetectable, sometimes with antiviral medicine during the study. Check what this study allows.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
HIV
The human immunodeficiency virus, which weakens the immune system.
Many studies now accept people whose HIV is controlled on treatment, judged by the viral load and the CD4 count. Check the limits this study sets.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Autoimmune disease
A condition in which the immune system attacks the body’s own tissues, such as rheumatoid arthritis, lupus, psoriasis, type 1 diabetes or inflammatory bowel disease.
Immunotherapy studies often leave out active autoimmune disease, and some name exceptions, such as thyroid disease on replacement hormone. Check the exceptions this study lists.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
QT and QTc
On an ECG, the QT interval is the time the heart’s lower chambers take to activate electrically and recover in each beat. QTc is that time adjusted for heart rate (QTcF by Fridericia’s formula, QTcB by Bazett’s). Some medicines lengthen it, and a long QTc can raise the risk of a dangerous heart rhythm.
The study may name a correction formula and a limit in milliseconds, often around 450–480.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
LVEF (left ventricular ejection fraction)
The share of blood the heart’s main pumping chamber (the left ventricle) pushes out with each beat, measured by an echocardiogram (heart ultrasound) or a MUGA scan. About 50–70% is normal.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
CTCAE grade
The US National Cancer Institute’s scale of how severe an adverse event is — any unwanted medical event during a study, such as a symptom, an illness or an abnormal test result, whether or not a treatment caused it. Whether it did is judged separately.
“Side effects of earlier treatment resolved to grade 1 or less” means they have settled to mild or gone.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Cancer (malignancy)
Malignancy means cancer. Criteria about another or earlier malignancy ask whether you have had a different cancer, usually within a set number of years.
Some are commonly allowed, such as treated skin cancers other than melanoma and early cancers removed completely — check the exceptions this study lists.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Hypersensitivity
An allergy, or another reaction of the immune system, to a substance. “Hypersensitivity to the study drug” means a known allergic reaction to it — not every side effect a medicine can have.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Control group will receive neither 5-fluorouracil (5FU) injection nor topical 0.005% calcipotriene cream. This group will receive standard of care only. Lesion will be surgical resected on day 84 of study.
5-fluorouracil (5FU) Group participants will receive a 1ml intralesional injection of 5FU 50mg/ml aqueous injectable solution. One injection will be administered once monthly for 3 month. Injections will occur on d0, d28, and d56. On the 4th visit the lesion site will be locally anesthetized and a 4mm punch biopsy will be taken to confirm pathologic resolution. Depending on the results of the confirmational biopsy the following actions will be taken: 1. Negative biopsy - clinical follow-up every 6 months for 2 years 2. Positive biopsy - Mohs surgery 2-4 weeks after
Drug: 5-fluorouracil
5-fluorouracil (5FU) + 0.005% calcipotriene cream Group participants will receive intralesional 5FU as in the previous group, additionally participants will also receive two (2) times daily for four (4) days one (1) day after each of three (3) 5FU injections topical application of 0.005% calcipotriene cream to the same lesion. On the 4th visit the lesion site will be locally anesthetized and a 4mm punch biopsy will be taken to confirm pathologic resolution. Depending on the results of the confirmational biopsy the following actions will be taken: 1. Negative biopsy - clinical follow-up every 6 months for 2 years 2. Positive biopsy - Mohs surgery 2-4 weeks after
Drug: 5-fluorouracil · Drug: Calcipotriene
Intralesional injections of 50mg/ml over a 3 week period.
Also known as: Adrucil, Tolak, Efudex, Carac, Fluoroplex, PremierPro Rx Fluorouracil, Fluorouracil Novaplus, 5FU
Topical application of .005% cream two (2) times daily for four (4) days one (1) day after each of three (3) 5FU injections..
Also known as: Dovonex, Calcitrene, Sorilux, Calsodore, Kalosar
Determine number of subjects experiencing Dose Limiting Toxicity (DLT) higher than grade 2, as defined by CTCAE v. 4.0
To assess Dose Limiting Toxicities (DLT) of intralesional-5FU and intralesional 5FU combined with topical imiquimod in subjects after 3 weeks of treatment. Dose Limiting Toxicities (DLT) are defined as: Higher-than grade 2 hematologic or non-hematologic toxicity that is definitely, probably, or possibly related to intralesional 5FU administration and/or topical imiquimod application. The NCI common terminology criteria for adverse events (CTCAE) version 4.0 will be used. Based on the results of the previous studies on treating SCCs with 5FU injection or topical imiquimod, significant toxicities are not expected. Adverse reactions were limited to local site reactions such as treatment site pain, induration, erythema, edema. If a patient has a DLT, doses will be delayed if any Grade \>2 toxicities are not resolved to Grade 1 by the time of the next dose.
Time frame: 84 days
Asses clinical objective response rate
Measure reduction in tumor burden to assess clinical objective response rate (ORR) in treated lesions. Response will be evaluated using modified Composite Assessment of Index Lesion Severity (CAILS) criteria by bi-dimensional measurement.
Time frame: 84 days
PDFs hosted by the registry, ClinicalTrials.gov.
Plan to share: Yes — Individual participant data that underlie the results reported in this article, after deidentification (text, tables, figures, and appendices)
Supporting information: Study protocol, Statistical analysis plan, Informed consent form, Clinical study report, Analytic code
Time frame: Immediately following publication. No end date.
Access criteria: Investigators whose proposed use of the data has been approved by an independent review committee (learned intermediary) identified for this purpose. To achieve aims in the approved proposal. Proposals may be submitted up to 36 months following article publication. Proposals should be directed to puglianomauroma@upmc.edu. To gain access, data requestors will need to sign a data access agreement. Data are available for 5 years through RedCap.
Data sharing details ↗ · www.ctsiredcap.pitt.edu
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Studies of the drugs tested here: Fluorouracil · Calcipotriene
Melissa Pugliano-Mauro