Perioperative Administration of COX 2 Inhibitors and Beta Blockers to Women Undergoing Breast Cancer Surgery
Perioperative Administration of COX 2 Inhibitors and Beta Blockers to Women Undergoing Breast Cancer Surgery
Surgery for breast cancer has a major role in enhancing long term survival and cure, but
 several physiological aspects associated with surgery are implicated as enhancing tumor
 spread and formation of distant metastases. These include: an increase in pro-angiogenic
 factors, direct spread of tumor cells, accumulation of grown factors, immune suppression and
 direct effects of anesthetics and opiate pain relievers on cancer cells. Some of these
 pro-metastatic mechanism may be blocked by the interventions proposed in this study, namely
 by administration of beta-adrenergic blockers and COX2 inhibitors around the time of surgery.

 Studies have shown that surgery increases levels of catecholamines and prostaglandins, which
 in turn may promote the release of pro-angiogenic factors such as VEGF, and enhance
 vascularization of micro metastases.

 Opiates given for pain relief during and after surgery have been reported to enhance tumor
 cell division and cause immune suppression.

 The immune system is significantly suppressed during surgery. This suppression has been shown
 to affect the systemic resistance to infection as well as neoplastic metastatic processes.

 Several studies have shown that increased levels of catecholamines and prostaglandins add to
 the immune suppression.

 Studies in rats found that peri-operative administration of the beta beta-blocker propranolol
 together with the COX2 inhibitor etodolac significantly reduced the suppression of NK cell
 activity as well as the risk for distant metastases.

 A recent retrospective clinical study found that among breast cancer patients treated with a
 combination of regional anesthesia and a COX inhibitor the recurrence rated were
 significantly less than among patients undergoing surgery without these two interventions.

 The purpose of the proposed prospective trial is to examine if peri-operative administration
 of the combination of a beta-blocker together with a COX2 inhibitor will prevent suppression
 of cellular immunity, decrease VEGF levels, and decrease cancer recurrence rates.

 In the proposed study breast cancer patients will be treated with a combination of a
 beta-blocker and COX2 inhibitor (or placebo) before, during and after surgery. (A control
 group of healthy women will serve as untreated controls). The variables which will be
 examined are: number and activity of NK cells, levels of Th1 and Th2 cytokines, serum stress
 hormones and angiogenic factors, and the ability of leukocytes to produce Th1 and Th2
 cytokines as a result of in vitro stimulation.

 In addition to these immediate parameters, long term follow up will be conducted in order to
 determine the effect of the intervention on long term cancer recurrence over five years.

 Statistical analysis will be done using t-tests, ANOVA, and multivariate regressions, with
 regard to the known risk factors for recurrence such as tumor grade, lymph node involvement
 etc. Sample size for immunological parameters will be 40 patients in each group and 20
 healthy women. Sample size for estimates of cancer recurrence at five years of follow up wiil
 be 460 women (230 in each group). This sample size provides a power of 80% to detect a 50%
 reduction in cancer recurrence at an α of 0.05.
Scientific Background Anesthesia and surgery are stressful situations which cause
 neuro-endocrine, metabolic and neurological responses. Cancer surgery may cause a decrease in
 anti-angiogenic factors, and enable dissemination of tumor cells through manipulation of the
 tumor and it blood vessels, as well as local and systemic secretion of growth hormones, and
 immune suppression. The magnitude of the response depends on many factors, including the
 extent of tissue injury, technique of anesthesia and type of analgesia, transfusions,
 temperature changes, psychological stress and genetic factors (1-3). The tissue damage starts
 a cascade of local and systemic processes which include secretion of hormones and cytokines
 (1).

 Post operatively, suppression of several immune functions occurs, and may last for weeks. The
 cellular immunity undergoes suppression, while the humoral immunity remains almost entirely
 intact (1). These changes in cell mediated immunity may impact systemic resistance to
 infection as well as metastatic processes. Specifically, NK cells have a pivotal role in
 defense from neoplastic processes. Studies have shown that high levels of activity of NK
 cells are associated with long term survival (1). Therefore, understanding of the processes
 which cause post-operative immune suppression, and prevention of such suppression, are of
 clinical significance.

 Several substances which are secreted after surgery are thought to contribute to immune
 suppression in general and NK cell dysfunction in particular. These include prostaglandins,
 catecholamines, steroids and endorphins, have all been shown to suppress several
 immunological functions in vitro (1). Animal studies have shown that administration of
 prostaglandins and catecholamines to rats suppresses NK cell activity in vivo, in association
 with elevated susceptibility to metastases 4-6).

 In rats undergoing a laparotomy, peri-operative administration of beta- adrenergic blockers
 together with COX-2 inhibitors significantly abrogated the NK cell suppression as well as the
 enhanced susceptibility to metastases after surgery (4,7).

 The mechanism of effect of prostaglandins and catecholamines on NK cells has been elucidated
 (1): beta- adrenergic receptors and prostanoid receptors on the cell membrane cause elevated
 levels of intra-cellular cAMP when activated by their ligand, which in turn interferes with
 the NK cell cytotoxic activity against cancer cells or viral pathogens. Since the elevated
 intra-cellular levels of cAMP is caused by each type of receptor independently, blockade of
 only one of these pathways will enable an increase in intracellular cAMP levels via the other
 pathway, and have no effect of the suppression of cytotoxic activity. Therefore, simultaneous
 blockade of both pathways may be more effective. Research in rats directly supports this
 hypothesis (7).

 Cells of the immune system express mostly beta-2 adrenergic receptors, and to a lesser degree
 beta-1 receptors (9, 10), so pharmacological blockade requires use of a non specific blocker
 which can block both types of receptors. The beta-antagonist propranolol was chosen for this
 experiment since it is a non specific beta-blocker, capable of blocking both types of
 receptors, as well as extensive clinical experience with this drug and the fact that it is a
 relatively safe drug. The COX-2 inhibitor chosen is Etodlac (Etopan), which has the advantage
 of being a selective COX-2 inhibitor,synthesized during injury and inflammation, with little
 effect on the COX-1 enzyme, which is associated with ongoing maintenance of tissues.

 Anesthetics and opiate analgesics also cause NK cell suppression. Research in animals and in
 humans has shown that morphine and fentanyl cause suppression of NK cell activity, and
 enhance susceptibility to breast cancer metastases in rats (11,12). Lower doses of opiates in
 patients have been reported to decrease the immune suppression magnitude and duration (13).
 Therefore, peri-operative interventions which decrease the use of opiates (or their
 endogenous secretion) may be clinically important factors in cancer recurrence.

 In addition to immune suppression, endogenous and exogenous opiates (endorphins morphine,
 fentanyl) have been implicated as promoting metastatic spread and neoplastic proliferation
 via other mechanisms:

 -  Catecholamines have been reported to increase VEGF secretion by human cancer cells, and
 enhance tumor cell invasion (14-16).

 -  Morphine causes increased proliferation of human tumor cells and enhances tumor
 vascularization (17)

 -  COX2 inhibitors enhance sensitivity of human tumor cells to induction of apoptosis, and
 decrease capillary density in cancer tissue (18, 19).

 Therefore, minimization of peri-operative opiate use and prevention or blockade of
 prostaglandin and catecholamine effects may delay the metastatic process and the development
 of existing micro-metastases.

 A retrospective clinical study published in 2006 indirectly supports the proposed study: In
 this study breast cancer patients were treated per-operatively with a COX inhibitor
 (diclofenac) along with local blockade of the sympathetic and pain pathways (paravertebral
 anesthesia). These treatments no doubt lowered the use of opiates during and after surgery.
 Among patients thus treated a three fold decrease in recurrence rates was found three years
 after surgery (21). Of note, this study quotes the previously mentioned study in rats (6,22)
 and involves the same interventions.

 AIM The aim of the proposed study is to examine whether peri-operative administration of COX2
 inhibitors with beta- adrenergic blockers decreases immune suppression during surgery,
 decreases stress hormone and pro-angiogenic factor secretion, and is associated with a
 decrease in cancer recurrence rates.

 The peri-operative variables which will be examined will include number and activity of NK
 cells in the blood, levels of Th1 and Th2 cytokines, stress hormones and pro-angiogenic
 factors, and Th1 and 2 cytokine production by leukocytes in response to in vitro stimulation.
 As part of the clinical monitoring meticulous recording of type and doses of analgesics given
 post operatively, as well as post operative pain assessment (NAS VAS). In addition to these
 parameters, long term follow up will be conducted to determine long term effects of the
 intervention on cancer recurrence.

 Peri-operative administration of COX inhibitors and beta- adrenergic blockers Routine pre
 medication before surgery generally includes anxiolytics such as benzodiazepines, drugs
 routinely taken by the patient, and additional drugs for specific indications. Preventive
 administration of beta- blockers and COX inhibitors are recommended in the literature
 pertaining to anesthesia and pain. Administration of beta- blockers has been shown to
 stabilize the cardio vascular system and decrease peri-operative ischemic events (23), while
 preventive administration of COX inhibitors decreases post operative pain (5). Post operative
 pain has significant immunological effects: it causes secretion of endogenous opiates, and is
 often treated with synthetic opiates such as morphine. The opiates cause a stress response,
 suppress immune functions and promote tumor growth and spread. Studies have shown that
 administration of COX inhibitors together with morphine caused a decrease in morphine
 requirements. Administration of COX inhibitors can help control pain by abrogation of
 prostaglandin mediated pain pathways while at the same time decreasing opiate requirements.
 An additional benefit of COX inhibitors is it's direct activity against tumor cells by
 enhancing apoptosis and decreasing tumor blood supply as reported in a recently published
 study (21). These studies indicate that peri-operative use of these drugs does not have any
 detrimental effects, is not associated with increased complication - and in fact may be
 beneficial.

 Patients and Methods

 Patients and treatments:

 Immune system activity will be determined among 80 operated breast cancer patients, and 20
 health women in the following groups:

 -  Treatment arm: 40 patients undergoing elective surgery for primary breast cancer will be
 treated orally according to the following protocol:

 -  Propranolol (Deralin) - patients will receive a low dose of 40 mg/day (4X10mg)
 starting two days before surgery (immediately following the first blood draw) until
 the evening of surgery. The first dose will be given under supervision (described
 later). On the morning of surgery, a single tablet of propranolol XL 80mg will be
 administered together with other pre-medication drugs, and will be continued once
 daily for three days after surgery. Pulse and blood pressure will be monitored
 during the first day of treatment, during the day of surgery and on the following
 days until discharge from the hospital. If pulse decreases to <50/min or blood
 pressure decreases to <100 systolic - the dose will be halved.

 -  Etodolac (Etopan) - patients will receive 400mg of etodolac XL twice a day starting
 two days before and continuing through two days after surgery.

 -  Control arm: Forty patients undergoing the same operations will be treated with placebo
 drugs on the same schedule.

 -  Healthy controls: a third control arm will include 20 healthy age-matched women
 presenting for routine breast exams and not undergoing surgery will undergo blood tests
 only.

 Patients will be randomized to one of the two groups, and patient, physicians and laboratory
 personnel will be blinded to the patient's assignment.

 Dependent variables to be tested:

 1. Cytotoxic activity of NK cells against K562 target cells.

 2. Blood leukocyte levels of NK cells, NKT cells, lymphocytes, monocytes and granulocytes
 as determined by FACS.

 3. Serum levels of cytokines (Th1, Th2 and inflammatory): IFN-gamma, IL-2, IL-12, IL-4,
 IL-10 IL-10 and IL-6.

 4. In vitro cytokine secretion by leukocytes stimulated by HPA and LPS. (IFN-gamma IL- 2,
 IL-12, IL-4, IL-10 IL-10 and IL-6.)

 5. Stress hormones and angiogenic factors: levels of cortisol and VEGF.

 6. Doses of anesthetics and analgesics used during and after surgery.

 7. Cancer recurrence during 5 years (including site of recurrence). In addition to the
 aforementioned groups, another 190 patients will be recruited onto each group without
 examining immunological parameters, and will be followed only for cancer recurrence.

 Eligibility criteria:

 -  Women that are scheduled to undergo surgery of a single, stage I-III, invasive ductal or
 lobular carcinoma tumor with curative intent

 -  No evidence of metastatic disease prior to surgery. Minimal workup would include chest
 XR, abdominal ultrasound and bone scan.

 -  Age between 20 and 70 year old.

 -  ASA score of 1-2

 -  The patient is able to understand the study objectives and procedures, able to comply
 with the protocol, and is capable to sign an informed consent.

 Exclusion criteria:

 -  Patients with metastatic disease, known prior to surgery.

 -  Patients in whom surgical resection is planned without curative intent.

 -  Patients who have undergone neoadjuvant treatment.

 -  Patients with renal failure, measured by creatinine level >1.5

 -  Patients with significant heart failure (NYHA functional class 3 or higher)

 -  Patients with significant liver failure (known cirrhosis, Bilirubin level>2)

 -  Patients suffering from asthma

 -  Patients with known allergy to one or more of the study medications

 -  Patients with known allergy to any medication from the non-steroidal anti-inflammatory
 drug group.

 -  Patients with diabetes mellitus (type 1 or 2).

 -  Patients treated chronically with one or more of the study medications

 -  Patients treated chronically with any type of Beta adrenergic blocker.

 -  Patients treated chronically with any type of COX inhibitor.

 -  Patients with second or third degree AV block.

 -  Patients with sinus bradycardia (patients with heart rate of less than 50).

 -  Patients with sick sinus syndrome.

 -  Patients with current atrial fibrillation/flutter.

 -  Patients with Printzmetal's angina

 -  Patients with hypertension

 -  Patients with right sided heart failure owing to pulmonary hypertension.

 -  Patients with significant cardiomegaly

 -  Patients with (current) pheochromocytoma

 -  Patients with chronic Digoxin treatment

 -  Patients with active peptic disease

 -  Patients with peripheral vascular disease

 -  Patients with history or concomitant malignant disease of any type other than breast
 cancer.

 -  Patients who were treated with chemotherapy in the last 10 years for any reason besides
 neo-adjuvant therapy for breast cancer within the last six months.

 -  Pregnant woman.

 -  Patients currently participating in any other clinical trial

 Procedure:

 After obtaining Helsinki committee approval, eligible patients will receive explanations and
 be invited to participate in the study. Women who consent will sign the approved informed
 consent form.

 Anesthesia procedure will include:

 Standard monitoring including automated blood pressure, ECG, O2 saturation, levels of inhaled
 and exhaled gases Pre-medication: at anesthesiologists discretion Induction: at
 anesthesiologist's discretion Maintenance: at anesthesiologist's discretion (an effort will
 be made to minimize opiate use but without compromising on pain control).

 Post operative pain management:

 Patients will be given a choice of oral Paracetamol 1000 mg every 4 hours, oral Dipyrone 1000
 mg every 4 hours, oral Tramadol solution 50 mg every 6 hours and combinations of the above.
 If needed - 5 mg of oral percocet/oxycodone syrup every 6 hours may be added. For patients
 who are unable to sustain oral intake, intravenous Tramal will be offered at a dose of 50-100
 mg every 6 hours and if needed - morphine 0.1 mg/kg every 4 hours. This protocol is identical
 to the one currently used after this type of surgery.

 Blood samples :

 Blood samples will be obtained two days before surgery (prior to administration of study
 drugs or placebo), on the morning of and the morning following surgery. Blood samples will be
 drawn between 7:30 and 9:00 am, and will include 10 cc of peripheral blood, preferably from
 the ante-cubital vein.

 Blood will be collected in vacuum tubes containing 30 units of heparin without preservatives,
 and will be transferred immediately to the neuro-immunology lab at Tel Aviv University.
 Laboratory evaluation of the samples will begin within 3 hours of blood drawing. The samples
 will be kept at room temperature.

 The samples will be divided as follows:

 NK cell activity testing - 2 ml FACS analysis of cells - 1 ml In vitro testing of cytokine
 response to LPS - 2 ml Cytokine, cortisol and VEGF levels - 5 ml (to be performed in batches
 at a later time - after separation of cells plasma will be kept at -80C).

 Documentation and analysis of results:

 Documentation:

 Epidemiological data: identification (name, ID no), age, ethnic origin, background medical
 conditions and drugs, smoking, menstrual status and date of last menstrual period, physical
 activity, presence of infections or viral illnesses within last 2 weeks.

 Data related to anesthesia and surgery:

 Type and length of surgery, systemic administration of opioids during and after surgery (type
 and dose).

 Cancer related parameters: Tumor size, grade, histological type, lymph node status, adjuvant
 therapy.

 Immunological parameters: as previously detailed is three blood samples from operated
 patients and a single sample from the healthy controls.

 Recurrence data: location, time since surgery and survival.

 Statistical Analysis:

 Statistical analysis will be done by ANOVA (between and within subjects) regarding the
 variables. Determination of specific differences between groups will be based on the PLSD
 test for planned comparisons, and the Scheffe test for unplanned comparisons. Chi square
 tests will be used as well as Kaplan-Meier survival curves for non continuous or categorical
 variables. In addition, Cox regressions will be calculated to evaluate the predictive ability
 of treatments on cancer recurrence, taking other predicting factors into account. Blood
 samples and immunological data will be collected for 40 patients in groups 1 and 2 and 20
 healthy controls in group 3. In order to determine impact of treatment on recurrence rates a
 total of 230 patients will be recruited in each group. Sample size was calculated to provide
 an 805 power to detect a 30-50% decrease in cancer recurrence with an alpha of 0.05.
Inclusion Criteria:

 -  Women that are scheduled to undergo surgery of a single, stage I-III, invasive ductal
 or lobular carcinoma tumor with curative intent

 -  No evidence of metastatic disease prior to surgery. Minimal workup would include chest
 XR, abdominal ultrasound and bone scan.

 -  Age between 20 and 70 year old.

 -  ASA score of 1-2

 -  The patient is able to understand the study objectives and procedures, able to comply
 with the protocol, and is capable to sign an informed consent.

 Exclusion Criteria:

 -  Patients with metastatic disease, known prior to surgery.

 -  Patients in whom surgical resection is planned without curative intent.

 -  Patients who have undergone neoadjuvant treatment.

 -  Patients with renal failure, measured by creatinine level >1.5

 -  Patients with significant heart failure (NYHA functional class 3 or higher)

 -  Patients with significant liver failure (known cirrhosis, Bilirubin level>2)

 -  Patients suffering from asthma

 -  Patients with known allergy to one or more of the study medications

 -  Patients with known allergy to any medication from the non-steroidal anti-inflammatory
 drug group.

 -  Patients with diabetes mellitus (type 1 or 2).

 -  Patients treated chronically with one or more of the study medications

 -  Patients treated chronically with any type of Beta adrenergic blocker.

 -  Patients treated chronically with any type of COX inhibitor.

 -  Patients with second or third degree AV block.

 -  Patients with sinus bradycardia (patients with heart rate of less than 50).

 -  Patients with sick sinus syndrome.

 -  Patients with current atrial fibrillation/flutter.

 -  Patients with Printzmetal's angina

 -  Patients with hypertension

 -  Patients with right sided heart failure owing to pulmonary hypertension.

 -  Patients with significant cardiomegaly

 -  Patients with (current) pheochromocytoma

 -  Patients with chronic Digoxin treatment

 -  Patients with active peptic disease

 -  Patients with peripheral vascular disease

 -  Patients with history or concomitant malignant disease of any type other than breast
 cancer.

 -  Patients who were treated with chemotherapy in the last 10 years for any reason
 besides neo-adjuvant therapy for breast cancer within the last six months.

 -  Pregnant woman.

 -  Patients currently participating in any other clinical trial